JP6703012B2 - Novel gadolinium chelate compounds for use in magnetic resonance imaging - Google Patents

Description

The invention is characterized by the claims, namely a novel high relaxivity extracellular gadolinium chelate based on a low molecular weight core polyamine, a method of preparing said compound, the use of said compound as an MRI contrast agent, And its use in the mammalian body.

1. Introduction The following nine gadolinium-based contrast agents (GBCA) are approved for clinical use: dimeglumine gadopentetate (Magnevist®), meglumine gadoterate (Dotarem®), gadoteridol (ProHance®). )), gadodiamide (Omniscan®), gadobutolol (Gadovist®), gadoversetamide (OptiMARK®), gadoxetic acid (Primovist®), dimeglumine gadobenate (MultiHance®). And gadophosbeset trisodium (Vasovist®/Ablavar®). With the exception of gadoxetate, dimeglumine gadobenate and gadophosbeset trisodium, GBCA exhibits a strictly extracellular, passive distribution in the body and is excreted exclusively via the kidney.

Gadoxetate and gadobenate dimeglumine show different pharmacokinetic profiles than other drugs. In addition to their extracellular distribution, they are taken up and also partially excreted via the liver. This is in addition to classical imaging possibilities (eg central nervous system, angiography, limb, heart, head/face/neck, abdomen and chest imaging) of GBCA on hepatocytes. It also allows imaging of the liver by the enhancement of liver parenchyma caused by uptake.

In contrast to other GBCAs, gadophosbeset trisodium does not show passive diffusion in the body and remains in the blood vessels. Reversible binding with HSA (human serum albumin) allows it to remain in the blood vessel for a long time, enabling high-resolution MR angiography.

The various GBCAs differ in their effects imparted by their longitudinal (r1) and transverse (r2) relaxivities, depending on magnetic field strength, temperature, and various intrinsic factors of metal chelates. The parameters that influence the intrinsic relaxivity are mainly the number of water molecules directly bound to gadolinium (so-called inner sphere water, q), the average residence time of inner sphere water molecules (τm), the second hydration layer. It is the number and residence time of water molecules and rotational diffusion (τr) in (so-called second sphere water) (Helm L. et al., Future Med Chem. 2010; 2:385-396). In terms of their relaxivity, all GBCAs on the market are very similar to each other and are obtained from the range of 4-7 L mmol ⁻¹ s ⁻¹ .

Strategies for increasing the sensitivity of GBCA are often described in the literature (Caravan P. et al., Chem. Soc. Rev., 2006, 35, 512-523, Helm et al., Future Med Chem. 2010; 2:385-. 396, Jacques V. Invest Radiol. 2010;45:613-624). One of the strategies is to increase the inner sphere water molecule (q), which is a water molecule directly coordinated to the gadolinium ion in the chelate. As the examples of AAZTA and HOPO based ligands show, increasing the inner sphere water molecules from one to two greatly increases relaxivity. Another strategy to increase relaxivity is to slow the rotational diffusion of molecules. The so-called rotational velocity (τr, see Introduction) describes the rotation of molecules in solution and is mainly influenced by the molecular size and protein binding of GBCA (Merbach AS et al., The Chemistry of Contrast Agents). in Medical Magnetic Resonance Imaging, 2013, ISBN:978-1-119-99176-2).

Another important feature of GBCAs is their complex stability. The potential for GBCA to release toxic free Gd ³⁺ ions is a major safety concern, especially for patients with end-stage renal disease. Renal systemic fibrosis (NSF) is a rare and serious syndrome associated with exposure to GBCA in patients with severe renal failure. NSF is associated with fibrotic changes in the skin and many organs. In 2010, the Food and Drug Administration (FDA) approved gadodiamide (Omniscan®), dimeglumine gadobenate (MultiHance®), dimeglumine gadopentetate (Magnevist®) and gadoversetamide (OptiMARK®). Published revised labeling recommendations for four GBCAs primarily associated with NSF, including: (Yang L et al., Radiology. 2012;265:248-253). At first glance, all GBCAs are very stable, but between linear and macrocyclic drugs, and between linear typical ionic and nonionic drugs. There is a big difference between them. Macrocycle GBCA has the highest complex stability (Frenzel T. et al., Invest Radiol. 2008;43:817-828). Increasing awareness of at-risk patients, the use of lower doses, and the further prevalence of macrocyclic GBCA have reduced the incidence of NSF over the last few years (Wang Y. et al. Radiology. 2011; 260:105-111 and Becker S. et al., Nephron Clin Pract. 2012;121:c91-c94).

A crucial issue in clinical applications is in vivo stability. Kinetic inactivity, together with thermodynamic stability, is the best predictor of in vivo toxicity of chelates with q=2, especially with regard to renal systemic fibrosis (NSF) risk (Merbach A. S. Et al., The Chemistry of Contrast Agents in Medical Magnetic Resonance Imaging, 2013, ISBN: 978-1-119-99176-2, p.157-208). The q=2 complex shows a two-fold improved relaxivity, but unfortunately is less stable than the q=1 compound (Hermann P. et al., Dalton Trans., 2008, 3027-3047).

2. Description of the Prior Art, Problems to be Solved and Their Solutions Several macrocyclic compounds have been described in the prior art.

EP1931673(B1) and EP2457914(B1) relate to pyDO3A (q=2) containing short amino alcohol chains, DO3A and DOTA compounds and metal complexes for medical imaging.

Macrocyclic lanthanide DO3A-like and DOTA-like GBCAs with high relaxivity have been described in the prior art.

Ranganathan R. S. Et al. (Investigative Radiology 1998; 33:779-797) investigated the effect of multimerization on the relaxivity of macrocyclic gadolinium chelates. WO 199531444 relates to monomeric and polymeric compounds with improved relaxivity.

No. 5,679,810 describes linear oligomeric polychelant compounds having alternating chelating and linker moieties linked by amide or ester moieties, and chelates formed therewith, and US Pat. Regarding its use in diagnostic imaging.

U.S. Pat. No. 5,650,133 discloses a bidentate chelator, in particular a compound having two macrocyclic chelator groups linked by a bridge containing ester or amide bonds, as well as its metal chelates and diagnostic imaging. Regarding its use in law.

WO 97/32862 (A1) a magnetic resonance imaging agent in which at least two units of a chelating agent are linked to the amino groups of the target carrier structure (eg protein, amino acid or peptide etc.). As a gadolinium polydentate chelating agent.

U.S. Patent Application Publication No. 2007/202047 relates to gadolinium chelate compounds for use in magnetic resonance imaging, which are 1,4,7,10-tetraazacyclododecane-1,4,7,10. -Derived from a chelating molecule selected from tetraacetic acid (DOTA) and diethylenetriaminepentaacetic acid (DTPA), at least one of the carboxylic acid groups of the chelating molecule reacts with an amine.

The more relaxed GBCA offers the opportunity to significantly reduce the dose on the one hand, while the standard dose is used to increase the sensitivity in MRI examinations of many diseases (Giesel FL. et al., Eur Radiol. 2010, 20:2461-2474).

European Patent No. 1931673 (B1) specification European Patent No. 2457914 (B1) International Publication No. 199531444 U.S. Pat.No. 5,679,810 U.S. Pat.No. 5650133 International Publication No. 97/32862 (A1) US Patent Application Publication No. 2007/202047

Helm L. Et al., Future Med Chem. 2010; 2:385-396 Caravan P. Chem. Soc. Rev. , 2006, 35, 512-523 Jacques V. Invest Radiol. 2010;45:613-624 Merbach A. S. Et al, The Chemistry of Contrast Agents in Medical Magnetic Resonance Imaging, 2013, ISBN: 978-1-119-99176-2. Yang L et al., Radiology. 2012;265:248-253 Frenzel T. Et al., Invest Radiol. 2008;43:817-828 Wang Y. Et al., Radiology. 2011;260:105-111 Becker S. Et al., Nephron Clin Pract. 2012;121:c91-c94 Hermann P. Dalton Trans. , 2008, 3027-3047 Investigative Radiology 1998;33:779-797. Giesel FL. Et al, Eur Radiol 2010, 20:2461-2474.

However, the medical need to provide GBCA for general use in magnetic resonance imaging such as the following has not been met:
Shows high relaxivity,
Exhibits an advantageous pharmacokinetic profile,
Completely excreted,
Chemically stable,
Shows high solubility in water,
Offers the potential for significant dose reduction,
It is very well tolerated, suitable for imaging various body regions.

Included in the above state of the art are certain high relaxivity extracellular gadolinium chelate compounds of general formula (I) of the invention as defined herein, or described and defined herein below. The stereoisomers, tautomers, N-oxides, hydrates, solvates or salts thereof referred to as "compounds of the invention" are not described.

It has now been found that the compounds according to the invention have surprisingly advantageous properties, which form the basis of the invention.

In particular, the compounds of the invention have a balanced high relaxivity profile, favorable pharmacokinetic profile, complete excretion, high stability, high solubility, potential for significant dose reduction and potential for systemic imaging. Has been demonstrated, and thus can be used as a contrast agent in magnetic resonance imaging (MRI).

The present invention describes a new class of high relaxivity extracellular gadolinium chelate complexes, methods for their preparation, and their use as MRI contrast agents.

According to a first aspect, the present invention provides four, five, six, seven or eight gadolinium [4,7,10-tris(carboxylatomethyl)-1,4,7,10- A general formula (I) containing a tetraazacyclododecan-1-yl] group
(In the formula:
Is:
and
(In the group, a and b each independently represent an integer of 1 or 2;
And,
In the group, * indicates the point of attachment of the group to R ^1. )
Represents a group selected from:
R ¹ is independently of each other a hydrogen atom or:
R ³ ,
and
(In the group, * indicates the point of attachment of the group to A)
Represents a group selected from
However, only ^one of the substituents R ¹ may represent a hydrogen atom;
n represents an integer of 3 or 4;
R ^2's each independently represent a hydrogen atom or a methyl group;
R ³ is:
and
(In the group, * indicates the point of attachment of the group to the rest of the molecule)
Represents a group selected from:
R ^4's each independently represent a hydrogen atom or a methyl group;
R ^5's each independently represent a hydrogen atom or a methyl group.)
Or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate or a salt thereof, or a mixture thereof.

The compounds of the present invention may contain one or more asymmetric centers, depending on the position and nature of the various substituents desired. Asymmetric carbon atoms may be present in the (R) or (S) configuration and may give rise to a racemic mixture if the asymmetric center is single and to a diastereomeric mixture if the asymmetric center is more than one. In certain instances, asymmetry may exist due to restricted rotation about a given bond, eg, the central bond connecting the two substituted aromatic rings of a particular compound.

Preferred compounds are those that produce the desired biological activity. Separated, pure or partially purified isomers and stereoisomers or racemic or diastereomeric mixtures of the compounds of this invention are also included within the scope of the present invention. Purification and separation of such materials can be accomplished by standard techniques known in the art.

Optical isomers can be obtained by resolution of racemic mixtures by conventional steps, eg, formation of diastereomeric salts with optically active acids or bases, or formation of covalent diastereomers. Examples of suitable acids are tartaric acid, diacetyl tartaric acid, ditoluoyl tartaric acid and camphorsulfonic acid. Mixtures of diastereoisomers may be separated into their individual diastereomers on the basis of their physical and/or chemical differences by methods known in the art, such as chromatography or fractional recrystallization. can do. The optically active base or acid is then liberated from the separated diastereomeric salt. Another step for separating optical isomers is to use the most appropriately chosen chiral chromatography (eg chiral HPLC column) to maximize the separation of enantiomers, with or without conventional derivatization. To do. Suitable chiral HPLC columns are manufactured by Daicel, for example Chiracel OD and Chiracel OJ are all routinely selectable, among others. Enzymatic separations are also useful with or without derivatization. The optically active compound of the present invention can also be obtained by chiral synthesis using optically active starting materials.

To identify the different types of isomers from each other, see IUPAC Rules Section E (Pure Appl Chem 45, 11-30, 1976).

The present invention relates to a single stereoisomer or as a mixture of said stereoisomers, for example R- or S-isomers in any ratio, or E- or Z-isomers. All possible stereoisomers of the compound are included. Isolation of a single stereoisomer of a compound of the invention, eg a single enantiomer or a single diastereomer, by any suitable state of the art method such as chromatography, especially chiral chromatography. May be realized.

Furthermore, the compounds of the present invention can exist as N-oxides, which is defined as at least one nitrogen of the compounds of the present invention being oxidized. The present invention includes all such possible N-oxides.

The present invention also relates to useful forms of the compounds disclosed herein, such as metabolites, hydrates, solvates, salts, especially pharmaceutically acceptable salts, and coprecipitates.

The compounds of the invention may exist as hydrates or solvates, wherein the compounds of the invention are polar solvents, in particular water, methanol or ethanol, for example the constitution of the crystal lattice of the compound. Include as an element. The amount of polar solvent, especially water, may be present in stoichiometric or non-stoichiometric ratio. In the case of stoichiometric solvates, for example, hydrates, hemi-, (semi-), mono-, sesqui-, di-, tri-, tetra-, penta-, etc. solvates or hydrates. Each thing is possible. The present invention includes all such hydrates or solvates.

Furthermore, the compounds of the present invention can exist in salt form. The salts may be either inorganic or organic addition salts, in particular any pharmaceutically acceptable inorganic or organic addition salts commonly used in pharmacy.

The term "pharmaceutically acceptable salt" refers to the relatively non-toxic, inorganic or organic acid addition salts of compounds of the present invention. For example, S. M. Berge et al., “Pharmaceutical Salts,” J. Pharm. Sci. See 1977, 66, 1-19. In particular, the production of neutral salts is described in US Pat. No. 5,560,903.

Pharmaceutically acceptable salts of the compounds according to the invention include salts of mineral acids and carboxylic acids, such as, but not limited to, hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, propionic acid, lactic acid, tartaric acid, malic acid. , Citric acid, fumaric acid, maleic acid, aspartic acid and glutamic acid salts.

Furthermore, one skilled in the art will appreciate that acid addition salts of the described compounds may be prepared by reacting the compounds with any suitable inorganic or organic acid via any of several known methods. Be understood.

The present invention includes all possible salts of the compounds of the present invention as a single salt or as a mixture of any of the above salts in any ratio.

In the present specification, especially in the experimental part, when the compounds are mentioned in the form of salts with the corresponding bases or acids for the synthesis of intermediates and examples of the invention, the respective preparation and/or purification steps The exact stoichiometric composition of the as-obtained salt form is in most cases unknown.

This means that, according to the described preparation and/or purification steps, the synthesis intermediates and example compounds or salts thereof as solvates of unknown stoichiometric composition, such as hydrates (if defined). The same applies to the case where is obtained.

According to a second embodiment of the first aspect, the present invention provides four, five or six gadolinium [4,7,10-tris(carboxylatomethyl)-1,4,7,10- The above general formula (I) containing a tetraazacyclododecan-1-yl] group (wherein:
Is:
and
(In the group, a and b each independently represent an integer of 1 or 2;
And,
In the group, * indicates the point of attachment of the group to R ^1. )
Represents a group selected from:
R ¹ is independently of each other a hydrogen atom or:
R ³ ,
and
(In the group, * indicates the point of attachment of the group to A)
Represents a group selected from
However, only ^one of the substituents R ¹ may represent a hydrogen atom;
n represents an integer of 3 or 4;
R ^2's each independently represent a hydrogen atom or a methyl group;
R ³ is:
and
(In the group, * indicates the point of attachment of the group to the rest of the molecule)
Represents a group selected from:
R ^4's each independently represent a hydrogen atom or a methyl group;
R ^5's each independently represent a hydrogen atom or a methyl group.)
Or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate or a salt thereof, or a mixture thereof.

According to a third embodiment of the first aspect, the present invention provides four, five or six gadolinium [4,7,10-tris(carboxylatomethyl)-1,4,7,10- The above general formula (I) containing a tetraazacyclododecan-1-yl] group (wherein:
Is:
and
(In the group, a and b represent an integer of 1;
And,
In the group, * indicates the point of attachment of the group to R ^1. )
Represents a group selected from:
R ¹ is independently of each other a hydrogen atom or:
R ³ ,
and
(In the group, * indicates the point of attachment of the group to A)
Represents a group selected from
However, only ^one of the substituents R ¹ may represent a hydrogen atom;
n represents an integer of 3 or 4;
R ² represents a hydrogen atom;
R ³ is:
and
(In the group, * indicates the point of attachment of the group to the rest of the molecule)
Represents a group selected from:
R ⁴ represents a hydrogen atom;
R ⁵ represents a hydrogen atom or a methyl group)
Or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate or a salt thereof, or a mixture thereof.

According to a fourth embodiment of the first aspect, the present invention provides four, five or six gadolinium [4,7,10-tris(carboxylatomethyl)-1,4,7,10- The above general formula (I) containing a tetraazacyclododecan-1-yl] group (wherein:
Is:
and
(In the group, a and b represent an integer of 1;
And,
In the group, * indicates the point of attachment of the group to R ^1. )
Represents a group selected from:
R ¹ is independently of each other a hydrogen atom or:
R ³ ,
and
(In the group, * indicates the point of attachment of the group to A)
Represents a group selected from
However, only ^one of the substituents R ¹ may represent a hydrogen atom;
n represents an integer of 3 or 4;
R ² represents a hydrogen atom;
R ³ is:
and
(In the group, * indicates the point of attachment of the group to the rest of the molecule)
Represents a group selected from:
R ⁴ represents a hydrogen atom;
R ⁵ represents a methyl group)
Or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate or a salt thereof, or a mixture thereof.

According to another aspect, the present invention provides a compound of general formula (I)
(In the formula:
Is
Represents a group (in the group, * represents the point of attachment of the group to R ¹ );
R ¹ represents the group R ³ ;
n represents an integer of 4;
R ² represents a hydrogen atom;
R ³ is:
and
(In the group, * indicates the point of attachment of the group to the rest of the molecule)
Represents a group selected from:
R ⁴ represents a hydrogen atom;
R ⁵ represents a hydrogen atom or a methyl group)
Or a stereoisomer, a tautomer, an N-oxide, a hydrate, a solvate or a salt thereof, or a mixture thereof.

In another embodiment of the above aspects, the invention provides 4, 5, 6, 7, or 8 gadolinium [4,7,10-tris(carboxylatomethyl)-1,4,7, A compound of formula (I) containing a 10-tetraazacyclododecan-1-yl] group.

In another embodiment of the above aspects, the invention provides 4, 5, or 6 gadolinium [4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane. A compound of formula (I) containing a -1-yl] group.

In another embodiment of the above aspects, the invention provides for four gadolinium [4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl] groups. To a compound of formula (I) comprising

In another embodiment of the above aspects, the invention provides five gadolinium [4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl] groups. To a compound of formula (I) comprising

In another embodiment of the above aspects, the invention provides 6 gadolinium [4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl] groups. To a compound of formula (I) comprising

In another embodiment of the above aspects, the invention provides 7 gadolinium [4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl] groups. To a compound of formula (I) comprising

In another embodiment of the above aspects, the invention provides eight gadolinium [4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl] groups. To a compound of formula (I) comprising

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
Is:
and
(In the group, a and b each independently represent an integer of 1 or 2;
And,
In the group, * represents a group selected from (indicates the point of attachment of the group to R ¹ )).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
Is:
and
(In the group, a and b each independently represent an integer of 1 or 2;
And,
In the group, * represents a group selected from (indicates the point of attachment of the group to R ¹ )).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
Is:
and
(In the group, a and b represent an integer of 1;
And,
In the group, * represents a group selected from (indicates the point of attachment of the group to R ¹ )).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
Is
Group (in the group, a and b each independently represent an integer of 1 or 2;
And,
In the groups, * represents a point of attachment of the group to R ¹ ).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
Is
Group (wherein a and b represent an integer of 1;
And,
In the groups, * represents a point of attachment of the group to R ¹ ).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
Is
A group (in the group, * represents a point of attachment of the group to R ¹ ).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
Is
Group (in the group, * indicates a point of attachment of the group to R ¹ ,
And
R ² represents a hydrogen atom))).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
Is
A group (in the group, * represents a point of attachment of the group to R ¹ ).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
Is
A compound representing a group (in the group, * represents a point of attachment of the group to R ¹ ).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
Is
A group (in the group, * represents a point of attachment of the group to R ¹ ).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
Is
A group (in the group, * represents a point of attachment of the group to R ¹ ).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
Is
A group (in the group, * represents a point of attachment of the group to R ¹ ).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
Is
A group (in the group, * represents a point of attachment of the group to R ¹ ).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ¹ is independently of each other a hydrogen atom or:
R ³ ,
and
(In the group, * indicates the point of attachment of the group to A,
Provided that only ^one of the substituents R ¹ may represent a hydrogen atom)).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ¹ is, independently of each other:
R ³ ,
and
(In the group, * represents a group selected from the point of attachment of the group to A)).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ¹ is, independently of each other:
R ³ and
(In the group, * represents a group selected from the point of attachment of the group to A)).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ¹ is, independently of each other:
R ³ and
(In the group, * represents a group selected from the point of attachment of the group to A)).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ¹ is, independently of each other:
R ³ and
(In the group, * represents a group selected from the point of attachment of the group to A)).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ¹ represents the group R ³ ).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ¹ is
A group (in the group, * represents the point of attachment of the group to A).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ¹ is
A group (in the group, * represents the point of attachment of the group to A).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ¹ is
A group (in the group, * represents the point of attachment of the group to A).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ^1's each independently represent a hydrogen atom or an R ³ group,
However, only ^one of the substituents R ¹ may represent a hydrogen atom).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ¹ is independently of each other a hydrogen atom or
Group (in the group, * indicates the point of attachment of the group to A)
Represents
However, it relates to a compound in which only ^one of the substituents R ¹ may represent a hydrogen atom.

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ¹ is independently of each other a hydrogen atom or
Group (in the group, * indicates the point of attachment of the group to A)
Represents
However, it relates to a compound in which only ^one of the substituents R ¹ may represent a hydrogen atom.

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ¹ is independently of each other a hydrogen atom or
Group (in the group, * indicates the point of attachment of the group to A)
Represents
However, it relates to a compound in which only ^one of the substituents R ¹ may represent a hydrogen atom.

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
n represents an integer of 3 or 4).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
n represents an integer of 3).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
n represents an integer of 4).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ² independently of each other represent a hydrogen atom or a methyl group).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ² represents a hydrogen atom).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ² represents a methyl group).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ³ is:
and
(In the group, * represents a group selected from the point of attachment of the group to the rest of the molecule).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ³ is
A group (wherein * represents the point of attachment of the group to the rest of the molecule).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ³ is
A group (wherein * represents the point of attachment of the group to the rest of the molecule).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ³ is
A group (in the group, * indicates the point of attachment of the group to the rest of the molecule);
And
R ⁵ represents a hydrogen atom or a methyl group).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ³ is
A group (in the group, * indicates the point of attachment of the group to the rest of the molecule);
And
R ⁵ represents a hydrogen atom).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ³ is
A group (in the group, * indicates the point of attachment of the group to the rest of the molecule);
And
R ⁵ represents a methyl group).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ⁴ independently of each other represent a hydrogen atom or a methyl group).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ⁴ represents a hydrogen atom).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ⁴ represents a methyl group).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ⁵ independently of each other represent a hydrogen atom or a methyl group).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ⁵ represents a hydrogen atom).

In another embodiment of the above aspects, the invention provides compounds of formula (I), wherein:
R ⁵ represents a methyl group).

It should also be understood that the present invention relates to any combination of the above embodiments.

Another embodiment of the first aspect is a compound of formula (I) selected from the group consisting of: or a stereoisomer, tautomer, N-oxide, hydrate, solvate or Salt or a mixture of them:
[4,10-bis(carboxylatomethyl)-7-{3,6,10,18,22,25-hexaoxo-26-[4,7,10-tris(carboxylatomethyl)-1,4,7 , 10-Tetraazacyclododecan-1-yl]-14-[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl] ] Propanoyl}amino)acetyl]-9,19-bis({[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl ] Propanoyl}amino)acetyl]amino}methyl)-4,7,11,14,17,21,24-heptaazaheptacosan-2-yl}-1,4,7,10-tetraazacyclododecane-1 -Ill] Acetate pentagadolinium,
[4,10-bis(carboxylatomethyl)-7-{3,6,10,15,19,22-hexaoxo-23-[4,7,10-tris(carboxylatomethyl)-1,4,7 ,10-Tetraazacyclododecan-1-yl]-9,16-bis({[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclo Dodecane-1-yl]propanoyl}amino)acetyl]amino}methyl)-11-(2-{[3-{[({2-[4,7,10-tris(carboxylatomethyl)-1,4, 7,10-Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}-2-({[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7 , 10-Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)propanoyl]amino}ethyl)-4,7,11,14,18,21-hexaazatetracosan-2-yl} -1,4,7,10-tetraazacyclododecan-1-yl]acetate hexagadolinium,
[4,10-bis(carboxylatomethyl)-7-{3,6,12,15-tetraoxo-16-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetra Azacyclododecan-1-yl]-9,9-bis({[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane-1- Ill]propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetraazaheptadecane-2-yl}-1,4,7,10-tetraazacyclododecan-1-yl]acetate tetra gadolinium,
{4,10-bis(carboxylatomethyl)-7-[(2R,16R)-3,6,12,15-tetraoxo-16-[4,7,10-tris(carboxylatomethyl)-1,4 ,7,10-Tetraazacyclododecan-1-yl]-9,9-bis({[({(2R)-2-[4,7,10-tris(carboxylatomethyl)-1,4,7 , 10-Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetraazaheptadecane-2-yl]-1,4,7,10-tetra Azacyclododecan-1-yl}acetate tetragadolinium,
{4,10-bis(carboxylatomethyl)-7-[(2S,16S)-3,6,12,15-tetraoxo-16-[4,7,10-tris(carboxylatomethyl)-1,4 ,7,10-Tetraazacyclododecan-1-yl]-9,9-bis({[({(2S)-2-[4,7,10-tris(carboxylatomethyl)-1,4,7 , 10-Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetraazaheptadecane-2-yl]-1,4,7,10-tetra Azacyclododecan-1-yl}acetate tetragadolinium,
[4-(1-{[2-(bis{2-[({1,4-bis[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7,10- Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]-1,4-diazepan-6-yl}carbonyl)amino]ethyl}amino)-2-oxoethyl]amino}-1-oxopropan-2-yl )-7,10-Bis(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetate pentagadolinium,
2, 2', 2'', 2''', 2'''', 2''''', 2'''''', 2''''''', 2'''''''',2''''''''',2'''''''''',2''''''''''',2'''''''''''',2''''''''''''',2'''''''''''''',2''''''''''''''',2'''''''''''''''',2'''''''''''''''''-{ethane-1,2-diylcarbamoyl-1 ,4-Diazepane-6,1,4-triyltris[(2-oxoethane-2,1-diyl)imino(1-oxopropane-1,2-diyl)-1,4,7,10-tetraazacyclododecane −10,1,4,7-Tetrayl]}octadecaacetate hexagadolinium,
2, 2', 2'', 2''', 2'''', 2''''', 2'''''', 2''''''', 2'''''''',2''''''''',2'''''''''',2''''''''''',2'''''''''''',2''''''''''''',2'''''''''''''',2''''''''''''''',2'''''''''''''''',2'''''''''''''''''-(1,4,7-triazonan-1,-1, 4,7-Triyltris{carbonyl-1,4-diazepan-6,1,4-triylbis[(2-oxoethane-2,1-diyl)imino(1-oxopropane-1,2-diyl)-1,4 , 7,10-Tetraazacyclododecane-10,1,4,7-tetrayl]}) octadecaacetate hexagadolinium,
2, 2', 2'', 2''', 2'''', 2''''', 2'''''', 2''''''', 2'''''''',2''''''''',2'''''''''',2'''''''''''-{1,4,7,10-tetra Azacyclododecane-1,4,7,10-tetrayltetrakis[(2-oxoethane-2,1-diyl)imino(1-oxopropane-1,2-diyl)-1,4,7,10-tetra Azacyclododecane-10,1,4,7-tetrayl]}dodecaacetate tetragadolinium,
2, 2', 2'', 2''', 2'''', 2''''', 2'''''', 2''''''', 2'''''''',2''''''''',2'''''''''',2''''''''''',2'''''''''''',2''''''''''''',2'''''''''''''',2''''''''''''''',2'''''''''''''''',2'''''''''''''''''-{3,7,10-triazatricyclo [3.3.3.0 ^1,5 ]undecane-3,7,10-triyltris[carbonyl (3,6,11,14-tetraoxo-4,7,10,13-tetraazahexadecane-8,2, 15-triyl)di-1,4,7,10-tetraazacyclododecane-10,1,4,7-tetrayl]}octadecaacetate hexagadolinium,
2, 2', 2'', 2''', 2'''', 2''''', 2'''''', 2''''''', 2'''''''',2''''''''',2'''''''''',2'''''''''''-(3,7,9-triazabicyclo [3.3.1] Nonane-3,7-diylbis{carbonyl-1,4-diazepan-6,1,4-triylbis[(2-oxoethane-2,1-diyl)-1,4,7,10 -Tetraazacyclododecane-10,1,4,7-tetrayl]}) dodecaacetate tetragadolinium,
{4,10-bis(carboxylatomethyl)-7-[2-oxo-2-({3-({[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetra Azacyclododecan-1-yl]acetyl}amino)-2,2-bis[({[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane-1- Ill]acetyl}amino)methyl]propyl}amino)ethyl]-1,4,7,10-tetraazacyclododecan-1-yl}acetate tetragadolinium, and [4,10-bis(carboxylatomethyl)-7] -{2,5,11,14-tetraoxo-15-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl]-8,8- Bis({[({[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetyl}amino)acetyl]amino}methyl)-3, 6,10,13-Tetraazapentadeca-1-yl}-1,4,7,10-tetraazacyclododecan-1-yl]acetate tetragadolinium.

According to another aspect, the present invention is directed to a method of preparing a compound of the invention, said method comprising the steps described in the experimental part herein.

According to another aspect, the present invention covers intermediate compounds useful for the preparation of the compounds of general formula (I) described above.

In particular, the invention has the general formula (II-a):
(In the formula,
Is as defined above for compounds of general formula (I), n'representing an integer of 2, 3 and 4) and salts thereof;
And the general formula (II-b):
(In the formula,
Is as defined above for compounds of general formula (I), n'representing an integer of 2, 3 and 4) and salts thereof;
And the general formula (II-c):
(In the formula,
Are as defined above for compounds of general formula (I), where n′ represents an integer of 2, 3 and 4) and its salts.

More particularly, the present invention is directed to intermediate compounds disclosed in the Examples section herein below.

According to another aspect, the present invention provides a compound of general formula (II-a) for the preparation of a compound of general formula (I) as defined above:
(In the formula,
Are as defined above for compounds of general formula (I), where n′ represents an integer of 2, 3 and 4) and are intended for use of the compounds and salts thereof.

According to another aspect, the present invention provides a compound of general formula (II-b) for the preparation of a compound of general formula (I) as defined above:
(In the formula,
Are as defined above for compounds of general formula (I), where n′ represents an integer of 2, 3 and 4) and are intended for use of the compounds and salts thereof.

According to another aspect, the present invention provides a compound of general formula (II-c) for the preparation of a compound of general formula (I) as defined above:
(In the formula,
Are as defined above for compounds of general formula (I), where n′ represents an integer of 2, 3 and 4) and are intended for use of the compounds and salts thereof.

According to another aspect, the present invention provides a compound of general formula (III) for the preparation of a compound of general formula (I) as defined above:
Where R ⁵ is as defined for the compound of general formula (I) above and LG is an activating leaving group such as 4-nitrophenol, or It covers the use of compounds of the formula (representing the groups defined for the synthesis of compounds of the formula (Ia)).

According to another aspect, the present invention provides a compound of general formula (IV) for the preparation of a compound of general formula (I) as defined above:
Where R ⁴ is as defined for the compound of general formula (I) above,
Represents a tetraamine as defined above for compounds of general formula (I)).

Another aspect of the invention is the use of compounds of general formula (I) for diagnostic imaging.

Preferably, the use of the compounds of the invention in diagnosis is carried out using magnetic resonance imaging (MRI).

Another aspect of the invention is a compound of general formula (I) for use in diagnostic imaging.

Another aspect of the invention is a compound of general formula (I) for use in magnetic resonance imaging (MRI).

The present invention also includes compounds of general formula (I) for the manufacture of diagnostic agents.

Another aspect of the invention is the use of a compound of general formula (I) or a mixture thereof for the manufacture of a diagnostic agent.

Another aspect of the invention is the use of a compound of general formula (I) or a mixture thereof for the manufacture of a diagnostic agent for magnetic resonance imaging (MRI).

Another aspect of the invention comprises the step of administering to a patient an effective amount of one or more compounds of general formula (I) in a pharmaceutically acceptable carrier, and performing NMR tomography on the patient. A method of imaging body tissue of a patient, including. Such a method is described in US Pat. No. 5,560,903.

For the manufacture of a diagnostic agent, eg for administration to a subject or test animal, a compound or mixture of general formula (I) will be conveniently formulated with a pharmaceutical carrier or excipient. The contrast agents of the present invention may conveniently include pharmaceutical formulation aids such as stabilizers, antioxidants, pH adjusters, fragrances and the like. Further, the production of the diagnostic medium according to the present invention is carried out by a method well known in the art (see US Pat. No. 5,560,903). They may be formulated for parenteral or enteral administration or for direct administration into body cavities. For example, parenteral formulations comprise a sterile solution or suspension of a compound of formula (I) according to the invention in a dose of 0.0001 to 5 mmol gadolinium/kg body weight, in particular 0.005 to 0.5 mmol gadolinium/kg body weight. .. Thus, the vehicle of the present invention may be in a physiologically acceptable carrier medium for injection, preferably in conventional pharmaceutical formulations such as solutions, suspensions, dispersions, syrups in water. When the contrast agent is formulated for parenteral administration, it is preferably isotonic or hypertonic, approaching pH 7.4.

In another aspect, the invention is directed to a method of diagnosing and monitoring the health of a patient. This method comprises the steps of: a) administering a compound of the invention to a human in need of such diagnosis to detect the compound in human, as described above and herein, and b) to the human. Measuring the signal generated by the administration of the compound of claim 1, preferably by magnetic resonance imaging (MRI).

General Synthesis Compounds according to the present invention can be prepared according to Schemes 1-12 below.

The schemes and procedures described below illustrate synthetic routes to the compounds of general formula (I) of the present invention and are not intended to be limiting. It will be apparent to those skilled in the art that the order of the transformations illustrated in the scheme can be changed in various ways. Therefore, the order of transformations illustrated in the scheme is not meant to be limiting. Suitable protecting groups and their introduction and cleavage are well known to those skilled in the art (e.g., T.W.Greene and P.G.M.Wuts in Protective Groups in Organic Synthesis, 3 rd edition, see Wiley 1999). Specific examples will be described in the following paragraphs.

The term "amine protecting group", as used herein, alone or as part of another group, is known or apparent to those skilled in the art and is a class of protecting groups: carbamate, amide, imide, Selected from, but not limited to, N-alkylamines, N-arylamines, imines, enamines, boranes, N-P protecting groups, N-sulphenyls, N-sulfonyls and N-silyls, and Textbooks Greene and Wuts, Protecting groups in Organic Synthesis, third edition, p. 494-653, but is not limited thereto. “Amine protecting group” is preferably carbobenzyloxy (Cbz), p-methoxybenzylcarbonyl (Moz or MeOZ), tert-butyloxycarbonyl (BOC), 9-fluorenylmethyloxycarbonyl (FMOC), benzyl. (Bn), p-methoxybenzyl (PMB), 3,4-dimethoxybenzyl (DMPM), p-methoxyphenyl (PMP), triphenylmethyl (Trityl), methoxyphenyldiphenylmethyl (MMT), or protected The amino group is a 1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl (phthalimido) or azido group.

The term "carboxyl protecting group," as used herein, alone or as part of another group, is known or apparent to those of skill in the art, and it is derived from a class of protecting groups: esters, amides and hydrazides. Selected, but not limited to, and described in the textbooks Greene and Wuts, Protecting groups in Organic Synthesis, third edition, p. 369-453, but is not limited thereto. The "carboxyl protecting group" is preferably methyl, ethyl, propyl, butyl, tert-butyl, allyl, benzyl, 4-methoxybenzyl or 4-methoxyphenyl.

The contents of the documents cited herein are hereby incorporated by reference.

A route for the preparation of compounds of general formula (Ia) is described in scheme 1.

FIG. 6 shows plasma kinetics of Example 3 versus Gadovist® in rats. The pharmacokinetic profile of Example 3 is comparable to that of Gadovist®. Example 3, Reference Compound 1 (Gadovist (R)), reference compound 2 (Magnevist (R)) and Reference Compound 3 (Primovist (TM)) paramagnetic relative water protons in the longitudinal relaxation rate R _{1 ^{p (t) / R 1 p}} (0) is a graph showing changes in versus time. The stability of Example 3 is comparable to the highly stable macrocyclic reference compound 1 (Gadovist®). FIG. 6 shows magnetic resonance angiography data in male New Zealand White rabbits. (A) 30 μmol Gd/kg bw Reference Compound 1 (Gadovist®), (B) 30 μmol Gd/kg bw Example 3, (C) 100 μmol Gd/kg bw Reference Compound 1. The contrast enhancement of the low dose protocol with Example 3 (B) is comparable to that of the standard dose of Reference Compound 1 (C). Moreover, the image quality of the low dose protocol of Example 3 (B) is far superior to that of the reference compound 1 (A). Angiographic studies show the possibility of a significant dose reduction in Example 3. It is an MR image before and after administration of a contrast agent. Representative images of the head and neck area before and 1.4 minutes after administration of Example 3 (A) and reference compound 1 (B). Strong signal enhancements are found, for example, in the heart, tongue and neck muscles. It is an MR image before and after administration of a contrast agent. Representative images of the abdomen before and 0.5 minutes after administration of Example 3 (A) and reference compound 1 (B). Strong signal enhancements are found, for example, in the aorta, kidney, liver and spleen. It is an MR image before and after administration of a contrast agent. Representative images of the pelvic area before and 2.9 minutes after administration of Example 3 (A) and reference compound 1 (B). Strong signal enhancements are found, for example, in the vascular system (blood vessels) and limb muscles. FIG. 6 is a diagram showing enhancement of MRI signals in various body regions. Enhancement of signal over time in tongue, oral muscle, liver, spleen, aorta and limb muscle after administration of Example 3 and reference compound 1 (Gadovist®). No difference in signal change over time was observed between Example 3 and Reference Compound 1. This shows the same pharmacokinetic properties, showing the potential of Example 3 in imaging different body regions. As expected from the about 2-fold relaxivity (see Example A), the observed contrast enhancement of Example 3 is greater than that of Reference Compound 1 (Gadovist®). Vertical bars represent standard deviation. It is a figure which shows the correlation of the gadolinium density|concentration of a tissue and the enhancement of MRI signal. After administration of Example 3 and Reference Compound 1, gadolinium concentration was measured in tissue samples of brain, tongue, liver, spleen, blood and limb muscle (muscle), and changes in MRI signals of each were determined in vivo. Vertical and horizontal error bars represent standard deviation. The dotted line represents a linear regression of gadolinium concentration and change in MRI signal. FIG. 6 shows the diffusion of various contrast agents through a semipermeable membrane (20 kDa). Dynamic CT measurements were performed to show the ability of various contrast agents to diffuse through the semipermeable membrane. (A) CT images of Examples 1, 2, 3, 4, 5 and 6 compared to reference compounds 1 (Gadovist®) and 4 (Gadomer). A typical measurement area in the evaluation of signals over time is shown in image A1. FIG. 6 shows signal analysis of a dynamic CT diffusion phantom survey over time. The Hounsfield Unit (HU) signal of the dialysis cassettes in Examples 1-6 and Reference Compounds 1 and 4 in fetal bovine serum solution was opposite to that of Reference Compound 4 (Gadomer) for all compounds investigated. Indicates that it can pass through a semipermeable membrane (20 kDa). Magnetic resonance imaging with enhanced contrast of rat GS9L brain tumors (indicated by white arrows). (A) Intra-individual comparison of Reference Compound 1 (Gadovist®) at the same 0.1 mmol Gd/kg body weight (bw) dose with Example 3. Example 3 showed higher lesion and brain contrast and excellent demarcation of the tumor margin. (B) Comparison of reference compound 1 (Gadovist®) at 0.3 mmol Gd/kg bw with Example 3 at 0.1 mmol Gd/kg bw. Example 3 showed similar lesion and brain contrast at one third of the reference compound 1 dose.

Scheme 1
Scheme 1: Route for the preparation of compounds of general formula (Ia). In the formula,
And R ⁵ have the meanings given for general formula (I) above, n′ represents an integer of 2, 3 and 4 and PG is for example a tert-butyloxycarbonyl group (BOC) Represents an amine protecting group such as or a group defined below.

Starting material 1 is a commercially available polyamine or a salt thereof [for example, CAS 111-40-0, CAS 28634-67-5, CAS 4730-54-5, CAS 4742-00-1, CAS 294-90-6], or It is either a polyamine or a salt thereof [eg CAS 41077-50-3] which is known from the literature or can be prepared analogously to the compounds described in the literature or in the experimental part below.

Triamine or tetraamine 1 or a salt thereof reacts with protected 3-amino-2-(aminomethyl)propionic acid 2-a [eg CAS 496974-25-5] or a salt thereof to give intermediate 3-a. Bring Suitable amine protecting groups for 3-amino-2-(aminomethyl)propionic acid are, for example, carbobenzyloxy (Cbz), p-methoxybenzylcarbonyl (Moz or MeOZ), tert-butyloxycarbonyl (BOC), 9 -Fluorenylmethyloxycarbonyl (FMOC), benzyl (Bn), p-methoxybenzyl (PMB), 3,4-dimethoxybenzyl (DMPM), p-methoxyphenyl (PMP), triphenylmethyl (Trityl), methoxy Phenyldiphenylmethyl (MMT), or the protected amino group is a 1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl (phthalimido) or azido group. The coupling reaction between the polyamine 1 and the propionic acid derivative 2-a can be performed, for example, in the presence of HATU and N,N-diisopropylethylamine in a suitable solvent (for example, N,N-dimethylformamide) at room temperature up to 80°C. Carried out using standard peptide coupling conditions, such as coupling within the temperature range up to, giving intermediates of general formula 3-a.

Deprotection of the intermediate of general formula 3-a resulting in the intermediate of general formula (II-a) or a salt thereof is described in the textbook Greene and Wuts, Protecting groups in Organic Synthesis, second edition, which is incorporated herein by reference. , P. It is carried out similarly to the method described in 309-405. The amine protecting group tert-butyloxycarbonyl (BOC) is a suitable solvent for the BOC protected intermediate of general formula 3-a (e.g. alcohol, tetrahydrofuran, dioxane or N,N-dimethylformamide, or mixtures thereof). ), and is removed by adding a suitable acid (such as an aqueous solution of hydrochloric acid or hydrobromic acid, or trifluoroacetic acid in an organic solvent such as dichloromethane). The deprotection reaction is carried out at a temperature ranging from room temperature to the boiling point of the respective solvent or solvent mixture, preferably the reaction is carried out at a temperature ranging from room temperature to 80°C.

The intermediate of the general formula (II-a) or a salt thereof has a leaving group (LG) (for example, pentafluorophenol, 4-nitrophenol, 1-hydroxypyrrolidine-2,5-dione, hydroxybenzotriazole or 3H-). [1,2,3]triazolo[4,5-b]pyridin-3-ol, etc.) to react the compound of general formula (Ia) with a Gd complex of general formula (III) Bring The preparation of activated esters is well known to those skilled in the art and is described, for example, by C.I. A. Montalbetti and V. Falque, Tetrahedron 61 (2005), p. 10827-10852. For example, 2,2′,2″-[10-(1-{[2-(4-nitrophenoxy)-2-oxoethyl]amino}-1-oxopropan-2-yl)-1,4,7 The preparation of ,10-tetraazacyclododecane-1,4,7-triyl]triacetato gadolinium is described in detail in WO2001051095(A2). The reaction of the intermediate of the general formula (II-a) with the activated Gd complex of the general formula (III) can be carried out in a suitable solvent such as dimethylsulfoxide, N,N-dimethylformamide, pyridine or mixtures thereof. ) And optionally the reaction is carried out in the presence of a base. Suitable bases are, for example, trialkylamines such as triethylamine or N,N-diisopropylethylamine. The reaction is carried out at a temperature ranging from room temperature to 100°C, preferably the reaction is carried out at a temperature ranging from 50°C to 70°C.

A route for the preparation of compounds of general formula (Ib) is described in scheme 2.

Scheme 2
Scheme 2: Route for the preparation of compounds of general formula (Ib). In the formula,
And R ⁵ have the meanings given for general formula (I) above, n′ represents an integer of 2, 3 and 4 and PG is for example a tert-butyloxycarbonyl group (BOC) Represents an amine protecting group such as or a group defined for the synthesis of compounds of general formula (Ia) above.

The compound of general formula (Ib) is synthesized in the same manner as the compound of general formula (Ia) as described above.

Starting material 1 is a commercially available polyamine or a salt thereof [for example, CAS 111-40-0, CAS 28634-67-5, CAS 4730-54-5, CAS 4742-00-1, CAS 294-90-6], or It is either a polyamine or a salt thereof [eg CAS 41077-50-3] which is known from the literature or can be prepared analogously to the compounds described in the literature or in the experimental part below.

Triamine or tetraamine 1 or a salt thereof reacts with protected 2,3-diaminopropionic acid 2-b [eg CAS 201472-68-6] or a salt thereof to give an intermediate of general formula 3-b, Gives the intermediate of the general formula (II-b) or a salt thereof after deprotection. In the final step, the intermediate of general formula (II-b) or salt thereof reacts with the Gd complex of general formula (III) to give a compound of general formula (I-b).

A route for the preparation of compounds of general formula (Ic) is described in scheme 3.

Scheme 3
Scheme 3: Route for the preparation of compounds of general formula (I-c). In the formula,
And R ⁵ have the meanings given for general formula (I) above, n′ represents an integer of 2, 3 and 4 and PG is for example a tert-butyloxycarbonyl group (BOC) Represents an amine protecting group such as or a group defined for the synthesis of compounds of general formula (Ia) above.

The compound of the general formula (Ic) is synthesized in the same manner as the compound of the general formula (Ia) as described above.

Starting material 1 is a commercially available polyamine or a salt thereof [for example, CAS 111-40-0, CAS 28634-67-5, CAS 4730-54-5, CAS 4742-00-1, CAS 294-90-6], or It is either a polyamine or a salt thereof [eg CAS 41077-50-3] which is known from the literature or can be prepared analogously to the compounds described in the literature or in the experimental part below.

Triamine or tetraamine 1 or salts thereof can be prepared as described in the experimental section below starting from methyl 1,4-dibenzyl-1,4-diazepan-6-carboxylate [see US Pat. No. 5,865,662]. Reaction with a protected 1,4-diazepan-6-carboxylic acid 2-c that can be synthesized affords an intermediate of the general formula 3-c which, after deprotection, has the general formula (II- The intermediate of c) or a salt thereof is provided. In the final step, the intermediate of general formula (II-c) or salt thereof reacts with the Gd complex of general formula (III) to give a compound of general formula (I-c).

A route for the preparation of compounds of general formula (Id) is described in scheme 4.

Scheme 4
Scheme 4: Route for the preparation of compounds of general formula (Id). In the formula,
R ⁵ has the meaning given for general formula (I) above,
Represents a tetraamine as described for general formula (I) above, LG is an activating leaving group such as 4-nitrophenol, or a compound of formula (Ia) above for synthesis. Represents a group defined for

Starting materials 4 are commercially available tetraamines or salts thereof [eg CAS 4742-00-1, CAS 294-90-6], or known from the literature or can be prepared analogously to compounds described in the literature. It is either tetraamine or a salt thereof.

Tetraamine 4 or a salt thereof is a leaving group (LG) (for example, pentafluorophenol, 4-nitrophenol, 1-hydroxypyrrolidine-2,5-dione, hydroxybenzotriazole or 3H-[1,2,3]triazolo). React with a Gd complex of general formula (III) activated by [4,5-b]pyridin-3-ol, etc.) to give a compound of general formula (Id). The preparation of activated esters is well known to those skilled in the art and is described, for example, by C.I. A. Montalbetti and V. Falque, Tetrahedron 61 (2005), p. 10827-10852. For example, 2,2′,2″-[10-(1-{[2-(4-nitrophenoxy)-2-oxoethyl]amino}-1-oxopropan-2-yl)-1,4,7 The preparation of ,10-tetraazacyclododecane-1,4,7-triyl]triacetato gadolinium is described in detail in WO2001051095(A2). The reaction of polyamine 4 or a salt thereof with the activated Gd complex of the general formula (III) is carried out in a suitable solvent such as dimethylsulfoxide, N,N-dimethylformamide, pyridine or a mixture thereof. Optionally, the reaction is carried out in the presence of a base. Suitable bases are, for example, trialkylamines such as triethylamine or N,N-diisopropylethylamine. The reaction is carried out at a temperature ranging from room temperature to 100°C, preferably the reaction is carried out at a temperature ranging from 50°C to 70°C.

A route for the preparation of compounds of general formula (Ie) is described in scheme 5.

Scheme 5
Scheme 5: Route for the preparation of compounds of general formula (Ie). In the formula,
R ⁴ has the meaning given for general formula (I) above,
Represents a tetraamine as described for general formula (I) above, LG is an activating leaving group such as 4-nitrophenol, or a compound of formula (Ia) above for synthesis. Represents a group defined for

Starting materials 4 are commercially available tetraamines or salts thereof [eg CAS 4742-00-1, CAS 294-90-6], or known from the literature or can be prepared analogously to compounds described in the literature. It is either tetraamine or a salt thereof.

Tetraamine 4 or a salt thereof has a leaving group (LG) (for example, pentafluorophenol, 4-nitrophenol, 1-hydroxypyrrolidine-2,5-dione [for example, tri-tert-butyl 2,2′,2′). '-(10-{2-[(2,5-dioxopyrrolidin-1-yl)oxy]-2-oxoethyl}-1,4,7,10-tetraazacyclododecane-1,4,7-triyl ) The synthesis of triacetate is described by Cong Li et al., J. Am. Chem. Soc. 2006, 128, p. 15072-15073; S3-5 and Galibert et al., Biorg. and Med. Chem. Letters 20 (2010), 5422-. 5425] or activated by hydroxybenzotriazole etc. [4,7,10-tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-]. Reaction with the tetraazacyclododecan-1-yl]acetic acid derivative 5 provides intermediate 6. The preparation of activated esters is well known to those skilled in the art and is described, for example, by C.I. A. Montalbetti and V. Falque, Tetrahedron 61 (2005), p. 10827-10852. The coupling reaction of polyamine 4 with [4,7,10-tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetic acid derivative 5 is It is carried out in a suitable solvent such as N,N-dimethylformamide or dimethylsulfoxide or a mixture thereof in the temperature range from room temperature up to 80° C. to give intermediate 6. Cleavage of the carboxyl protecting group of intermediate 6 to give the intermediate of general formula (IV) is described in the textbook Greene and Wuts, Protecting groups in Organic Synthesis, second edition, p. It can be realized as described in 245-247. Deprotection is carried out, for example, by dissolving and stirring Intermediate 6 in trifluoroacetic acid at room temperature for several hours. Complexation of an intermediate of general formula (IV) with a suitable gadolinium(III) compound or salt, such as gadolinium oxide, gadolinium triacetate or gadolinium triacetate hydrate, gadolinium trichloride or gadolinium trinitrate. Are well known to those skilled in the art. The intermediate of general formula (IV) is dissolved in water and after addition of the appropriate gadolinium(III) compound, the resulting mixture is stirred for several hours at pH=1-7 within the temperature range from room temperature up to 100°C. To give compounds of general formula (Ie). The intermediate of the general formula (IV) is, for example, dissolved in water, gadolinium triacetate tetrahydrate is added, and a suitable base (for example, an aqueous sodium hydroxide solution) is added to adjust the pH to 3.5-. Adjusted to 5.5. The reaction is carried out at a temperature in the range of 50° C. to 80° C., leading to compounds of general formula (Ie).

A route for the preparation of compounds of general formula (If) is described in scheme 6.

Scheme 6
Scheme 6: Route for the preparation of compounds of general formula (If). In the formula,
Is a triamine as defined above, n′ represents an integer of 2, or
Is a tetraamine as defined above, n'represents an integer of 3,
R ⁵ has the meaning given for general formula (I) above, LG represents an activated leaving group, for example 4-nitrophenol, or a group as defined below.

The intermediate of the general formula (II-a) or a salt thereof described in Scheme 1 and the experimental part below (wherein n represents an integer of 2,
Represents a triamine core as defined above. ), or an intermediate of the general formula (II-a) or a salt thereof (wherein n′ represents an integer of 3,
Represents a tetraamine core as defined above. ) Is a leaving group (LG) (eg, pentafluorophenol, 4-nitrophenol, 1-hydroxypyrrolidine-2,5-dione, hydroxybenzotriazole or 3H-[1,2,3]triazolo[4,5] -B] pyridin-3-ol and the like) reacts with a Gd complex of general formula (III) to give a compound of general formula (If). The preparation of activated esters is well known to those skilled in the art and is described, for example, by C.I. A. Montalbetti and V. Falque, Tetrahedron 61 (2005), p. 10827-10852. For example, 2,2′,2″-[10-(1-{[2-(4-nitrophenoxy)-2-oxoethyl]amino}-1-oxopropan-2-yl)-1,4,7 The preparation of ,10-tetraazacyclododecane-1,4,7-triyl]triacetato gadolinium is described in detail in WO2001051095(A2). The reaction of the intermediate of the general formula (II-a) or a salt thereof with the activated Gd complex of the general formula (III) can be carried out in a suitable solvent (for example, dimethylsulfoxide, N,N-dimethylformamide, pyridine or the like). , Etc.) and optionally the reaction is carried out in the presence of a base. Suitable bases are, for example, trialkylamines such as triethylamine or N,N-diisopropylethylamine. The reaction is carried out at a temperature ranging from room temperature to 100°C, preferably the reaction is carried out at a temperature ranging from 50°C to 70°C.

A route for the preparation of compounds of general formula (Ig) is described in scheme 7.

Scheme 7
Scheme 7: Route for the preparation of compounds of general formula (Ig). In the formula,
Is a triamine as defined above, n′ represents an integer of 2, or
Is a tetraamine as defined above, n′ represents an integer of 3, R ⁵ has the meaning given for general formula (I) above and LG is for example 4-nitro. Represents an activated leaving group such as phenol, or a group defined below.

The compound of the general formula (Ig) is synthesized in the same manner as the compound of the general formula (If) as described above.

An intermediate of the general formula (II-b) described in Scheme 2 or a salt thereof (in the formula, n′ represents an integer of 2,
Represents a triamine core as defined above. ), or an intermediate of the general formula (II-b) or a salt thereof (in the formula, n′ represents an integer of 3,
Represents a tetraamine core as defined above. ) Is a leaving group (LG) (eg, pentafluorophenol, 4-nitrophenol, 1-hydroxypyrrolidine-2,5-dione, hydroxybenzotriazole or 3H-[1,2,3]triazolo[4,5] -B] pyridin-3-ol and the like) reacts with a Gd complex of general formula (III) to give a compound of general formula (I-g).

A route for the preparation of compounds of general formula (Ih) is described in scheme 8.

Scheme 8
Scheme 8: Route for the preparation of compounds of general formula (I-h). In the formula,
Is a triamine as defined above, n′ represents an integer of 2, or
Is a tetraamine as defined above, n′ represents an integer of 3, R ⁵ has the meaning given for general formula (I) above and LG is for example 4-nitro. Represents an activated leaving group such as phenol, or a group defined below.

The compound of the general formula (Ih) is synthesized in the same manner as the compound of the general formula (If) as described above.

An intermediate of the general formula (II-c) described in Scheme 3 or a salt thereof (in the formula, n′ represents an integer of 2,
Represents a triamine core as defined above. ), or an intermediate of the general formula (II-c) or a salt thereof (in the formula, n′ represents an integer of 3,
Represents a tetraamine core as defined above. ) Is a leaving group (LG) (eg pentafluorophenol, 4-nitrophenol, 1-hydroxypyrrolidine-2,5-dione, hydroxybenzotriazole or 3H-[1,2,3]triazolo[4,5] -B]pyridin-3-ol, etc.) reacts with a Gd complex of general formula (III) to give a compound of general formula (Ih).

A route for the preparation of compounds of general formula (Ik) is described in scheme 9.

Scheme 9
Scheme 9: Route for the preparation of compounds of general formula (Ik). In the formula,
And R ⁴ have the meanings given for general formula (I) above, n′ represents an integer of 2, 3 and 4 and LG represents for example 1-hydroxypyrrolidine-2,5- Represents an activated leaving group such as a dione, or a group as defined for the synthesis of compounds of general formula (Ia) above, PG represents a carboxyl protecting group such as a methyl or ethyl group.

Starting material 1 is a commercially available polyamine or a salt thereof [for example, CAS 111-40-0, CAS 28634-67-5, CAS 4730-54-5, CAS 4742-00-1, CAS 294-90-6], or It is either a polyamine or a salt thereof [eg CAS 41077-50-3] which is known from the literature or can be prepared analogously to the compounds described in the literature or in the experimental part below.

Diamine 7 or a salt thereof is commercially available [for example, CAS 1417898-94-2], or can be synthesized by a method well known to those skilled in the art. The diamine 7 or a salt thereof has a leaving group (LG) (eg, pentafluorophenol, 4-nitrophenol, 1-hydroxypyrrolidine-2,5-dione [eg, tri-tert-butyl 2,2′,2′). '-(10-{2-[(2,5-dioxopyrrolidin-1-yl)oxy]-2-oxoethyl}-1,4,7,10-tetraazacyclododecane-1,4,7-triyl ) The synthesis of triacetate is described by Cong Li et al., J. Am. Chem. Soc. 2006, 128, p. 15072-15073; S3-5 and Galibert et al., Biorg. and Med. Chem. Letters 2010, 20, p. 5422. -5425] or activated by hydroxybenzotriazole, etc. [4,7,10-tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10- It can be reacted with the tetraazacyclododecan-1-yl]acetic acid derivative 5 to give the intermediate 8. The preparation of activated esters is well known to those skilled in the art and is described, for example, by C.I. A. Montalbetti and V. Falque, Tetrahedron 2005, 61 p. 10827-10852. The protective group PG of the intermediate 8 is cleaved to give a carboxylic acid by cleaving in water or a mixture of water and tetrahydrofuran under basic conditions, for example, by treatment with an alkali metal hydroxide (such as lithium hydroxide). Corresponding salt can be given. This salt is prepared, for example, in the presence of HATU and 3H-[1,2,3]triazolo[4,5-b]pyridin-3-ol in the presence of N,N-diisopropylethylamine in a suitable solvent (eg dichloromethane). Can be coupled with polyamine 1 using standard peptide coupling conditions, such as coupling at room temperature in) to give intermediates of general formula (V). Cleavage of the carboxyl protecting group of the intermediate of general formula (V) can be achieved using standard conditions, for example by dissolving and stirring intermediate (V) in aqueous hydrochloric acid at room temperature. Subsequent complexation with a suitable gadolinium(III) compound or salt, such as gadolinium oxide, gadolinium triacetate or gadolinium triacetate hydrate, gadolinium trichloride or gadolinium trinitrate, etc., is well known to those skilled in the art, For example, it can be realized by reacting with a suitable gadolinium (III) compound for several hours at a pH of 1 to 7 within a temperature range from room temperature to a maximum of 100° C. to give a compound of the general formula (Ik). it can. Untreated carboxylic acids derived from compounds of general formula (V) react with, for example, gadolinium oxide at 80° C. to give compounds of general formula (Ik).

A route for the preparation of compounds of general formula (I-m) and (I-n) is described in scheme 10.

Scheme 10
Scheme 10: Route for the preparation of compounds of general formula (I-m) and (I-n). In the formula,
And R ⁴ have the meanings given for general formula (I) above, n′ represents an integer of 2, 3 and 4 and LG represents for example 1-hydroxypyrrolidine-2,5- Represents an activated leaving group such as a dione, or a group as defined for the synthesis of compounds of general formula (Ia) above, PG represents a carboxyl protecting group such as a methyl or ethyl group.

Instead of the diamine of formula 7 described in scheme 9, when the diamine of formula 9 and 10 or salts thereof is used in a similar synthesis as described in scheme 9, the general formula (I-m) and The compound (I-n) can be obtained.

Diamine 9 or a salt thereof is commercially available [for example, CAS 159029-33-1 and CAS 440644-06-4], or can be synthesized by a method well known to those skilled in the art.

Diamine 10 or a salt thereof is commercially available [for example, CAS 20610-20-2, CAS 6059-44-5], or can be synthesized by a method well known to those skilled in the art.

An alternative route to the route described in Scheme 4 for the preparation of compounds of general formula (Id) is described in Scheme 11.

Scheme 11
Scheme 11: Alternative route for the preparation of compounds of general formula (Id). In the formula,
R ⁵ has the meaning given for general formula (I) above,
Represents a tetraamine as described for general formula (I) above, and LG represents an activated deprotection such as 3H-[1,2,3]triazolo[4,5-b]pyridin-3-ol. Represents a leaving group or a group as defined for the synthesis of the compounds of general formula (Ia) above.

Starting materials 4 are commercially available tetraamines or salts thereof [eg CAS 4742-00-1, CAS 294-90-6], or known from the literature or can be prepared analogously to compounds described in the literature. It is either tetraamine or a salt thereof. Starting materials 14 are either commercially available or known from the literature or can be synthesized analogously to the compounds described in the literature, for example by stepwise alkylation of a cyclen core.

Tetraamine 4 or a salt thereof has a leaving group (LG) (for example, 1-hydroxypyrrolidine-2,5-dione, pentafluorophenol, 4-nitrophenol or 3H-[1,2,3]triazolo[4,5] -B] pyridin-3-ol, etc.) to react with amino acid derivative 11 to give intermediate 12. The preparation of activated esters is well known to those skilled in the art and is described, for example, by C.I. A. Montalbetti and V. Falque, Tetrahedron 61 (2005), p. 10827-10852. The coupling reaction between polyamine 4 and amino acid derivative 11 is carried out in a suitable solvent (eg dichloromethane or N,N-dimethylformamide etc.) within a temperature range from room temperature up to 50° C. to give intermediate 12. .. Cleavage of the amino protecting group (PG) of intermediate 12 to give intermediate 13 can be accomplished as described in the textbook Greene and Wuts, Protecting groups in Organic Synthesis, second edition. In the case of a tert-butoxycarbonyl protecting group, deprotection can be carried out, for example, in a suitable solvent such as CPME or 1,4-dioxane, or mixtures thereof, in the temperature range from 0° C. to room temperature, the intermediate 12 Is carried out by reacting with HCl in CPME for several hours.

Tetraamine 13 or a salt thereof has a leaving group (LG) (eg, 3H-[1,2,3]triazolo[4,5-b]pyridin-3-ol, 4-nitrophenol or 1-hydroxypyrrolidine-2). ,5-dione, etc.) activated [4,7,10-tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetic acid derivative Reacts with 14 to give intermediate 15. The coupling reaction of tetraamine 13 with [4,7,10-tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetic acid derivative 14 is It is carried out in a suitable solvent such as N,N-dimethylacetamide or dimethylsulfoxide or a mixture thereof in the temperature range from room temperature to 80° C. to give intermediate 15.

Cleavage of the carboxyl protecting group of intermediate 15, which gives intermediates of general formula (VI), is described in textbooks Greene and Wuts, Protecting groups in Organic Synthesis, second edition, p. It can be realized as described in 245-247. Deprotection is carried out, for example, by dissolving and stirring Intermediate 15 in trifluoroacetic acid at room temperature for several hours.

Complexation of an intermediate of general formula (VI) with a suitable gadolinium(III) compound or salt, such as gadolinium oxide, gadolinium triacetate or gadolinium triacetate hydrate, gadolinium trichloride or gadolinium trinitrate. Are well known to those skilled in the art. The intermediate of general formula (VI) is dissolved in water and after addition of a suitable gadolinium(III) compound, the resulting mixture is stirred for a few hours at pH=1-7 within a temperature range from room temperature up to 100°C. To give compounds of general formula (Id). The intermediate of the general formula (VI) is, for example, dissolved in water, gadolinium triacetate tetrahydrate is added, and a suitable base (for example, an aqueous sodium hydroxide solution) is added to adjust the pH to 3.5 to. Adjusted to 5.5. The reaction is carried out at a temperature in the range of 50° C. to 80° C., leading to compounds of general formula (Id).

An alternative route to the route described in Scheme 4 for the preparation of compounds of general formula (Id) is described in Scheme 12.

Scheme 12
Scheme 12: Alternative route for the preparation of compounds of general formula (Id). In the formula,
R ⁵ has the meaning given for general formula (I) above,
Represents a tetraamine as described for general formula (I) above, and LG represents an activated deprotection such as 3H-[1,2,3]triazolo[4,5-b]pyridin-3-ol. Represents a leaving group or a group as defined for the synthesis of the compounds of general formula (Ia) above.

Starting materials 4 are commercially available tetraamines or salts thereof [eg CAS 4742-00-1, CAS 294-90-6], or known from the literature or can be prepared analogously to compounds described in the literature. It is either tetraamine or a salt thereof. The starting material 16 is known from the literature or can be synthesized analogously to the compounds described in the literature, for example by stepwise alkylation of the cyclen core.

Tetraamine 4 or a salt thereof has a leaving group (LG) (for example, 3H-[1,2,3]triazolo[4,5-b]pyridin-3-ol, 4-nitrophenol or 1-hydroxypyrrolidine-2). ,5-dione, etc.) activated [4,7,10-tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetic acid derivative Reacts with 16 to give intermediate 15. The coupling reaction between tetraamine 4 and [4,7,10-tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetic acid derivative 16 is Performed in a suitable solvent such as N,N-dimethylformamide to provide intermediate 16.

Complexation of an intermediate of general formula (VI) with a suitable gadolinium(III) compound or salt, such as gadolinium oxide, gadolinium triacetate or gadolinium triacetate hydrate, gadolinium trichloride or gadolinium trinitrate. Are well known to those skilled in the art. The intermediate of general formula (VI) is dissolved in water and after addition of a suitable gadolinium(III) compound, the resulting mixture is stirred for a few hours at pH=1-7 within a temperature range from room temperature up to 100°C. To give compounds of general formula (Id). The intermediate of the general formula (VI) is, for example, dissolved in water, gadolinium triacetate tetrahydrate is added, and a suitable base (for example, an aqueous sodium hydroxide solution) is added to adjust the pH to 3.5 to. Adjusted to 5.5. The reaction is carried out at a temperature in the range of 50° C. to 80° C., leading to compounds of general formula (Id).

According to certain embodiments, the present invention also relates to a method of preparing a compound of general formula (I-a) as defined above, said method comprising general formula (II-a):
(In the formula,
Is as defined for the compound of general formula (I) above, and n′ represents an integer of 2, 3 and 4) or an intermediate compound thereof,
General formula (III):
Where R ⁵ is as defined for the compound of general formula (I) above, and LG is an activated leaving group such as 4-nitrophenol, or the general formula (I- a) represents a group defined for the synthesis of compounds)
Of a compound of general formula (Ia):
(In the formula,
And R ⁵ is as defined for compounds of general formula (I) above, and n′ represents an integer of 2, 3 and 4).

According to another embodiment, the present invention also relates to a method of preparing a compound of general formula (I-b) as defined above, said method comprising general formula (II-b):
(In the formula,
Is as defined for the compound of general formula (I) above, and n′ represents an integer of 2, 3 and 4) or an intermediate compound thereof,
General formula (III):
Where R ⁵ is as defined for the compound of general formula (I) above, and LG is an activated leaving group such as 4-nitrophenol, or the general formula (I- a) represents a group defined for the synthesis of compounds)
Of a compound of general formula (Ib):
(In the formula,
And R ⁵ is as defined for compounds of general formula (I) above, and n′ represents an integer of 2, 3 and 4).

According to another embodiment, the present invention also relates to a method of preparing a compound of general formula (I-c) as defined above, said method comprising general formula (II-c):
(In the formula,
Is as defined for the compound of general formula (I) above, and n′ represents an integer of 2, 3 and 4) or an intermediate compound thereof,
General formula (III):
Where R ⁵ is as defined for the compound of general formula (I) above, and LG is an activated leaving group such as 4-nitrophenol, or the general formula (I- a) represents a group defined for the synthesis of compounds)
Of a compound of general formula (I-c):
(In the formula,
And R ⁵ is as defined for compounds of general formula (I) above, and n′ represents an integer of 2, 3 and 4).

According to another embodiment, the present invention also relates to a method of preparing a compound of general formula (Id) as defined above, said method comprising:
(In the formula,
Is a tetraamine as defined above for compounds of general formula (I)) or a salt thereof,
General formula (III):
Where R ⁵ is as defined for the compound of general formula (I) above, and LG is an activated leaving group such as 4-nitrophenol, or the general formula (I- a) represents a group defined for the synthesis of compounds)
Of a compound of general formula (Id):
Where R ⁵ is as defined for the compound of general formula (I) above,
Includes a compound of formula (I) which is a tetraamine as defined above for compounds of general formula (I).

According to another embodiment, the present invention also relates to a method of preparing a compound of general formula (Ie) as defined above, said method comprising general formula (IV):
Where R ⁴ is as defined for the compound of general formula (I) above,
Is a tetraamine as defined above for compounds of general formula (I)),
A gadolinium (III) compound (eg, gadolinium oxide, gadolinium triacetate or gadolinium triacetate hydrate, gadolinium trichloride or gadolinium trinitrate) or a salt thereof,
Thereby, the general formula (Ie):
Where R ⁴ is as defined for the compound of general formula (I) above,
Includes a compound of formula (I) which is a tetraamine as defined above for compounds of general formula (I).

According to another embodiment, the present invention also relates to a method of preparing a compound of general formula (If) as defined above, said method comprising general formula (II-a):
(In the formula,
Is a triamine as defined above for compounds of general formula (I), n'represents an integer of 2, or, in the formula:
Is a tetraamine as defined for the compound of general formula (I) above, and n′ represents an integer of 3) or an intermediate compound thereof,
General formula (III):
Where R ⁵ is as defined for the compound of general formula (I) above, and LG is an activated leaving group such as 4-nitrophenol, or the general formula (I- a) represents a group defined for the synthesis of compounds)
Of a compound of general formula (I-f):
Where R ⁵ is as defined for the compound of general formula (I) above, wherein
Is a triamine as defined above for compounds of general formula (I), n'represents an integer of 2, or, in the formula:
Is a tetraamine as defined above for compounds of general formula (I), n'representing an integer of 3).

According to another embodiment, the present invention also relates to a method of preparing a compound of general formula (Ih) as defined above, said method comprising general formula (II-c):
(In the formula,
Is a triamine as defined above for compounds of general formula (I), n'represents an integer of 2, or, in the formula:
Is tetraamine as defined above for compounds of general formula (I), n'representing an integer of 3)
Of the intermediate compound or a salt thereof,
General formula (III):
Where R ⁵ is as defined for the compound of general formula (I) above, and LG is an activated leaving group such as 4-nitrophenol, or the general formula (I- a) represents a group defined for the synthesis of compounds)
Of a compound of general formula (I-h):
Where R ⁵ is as defined for the compound of general formula (I) above, wherein
Is a triamine as defined above for compounds of general formula (I), n'represents an integer of 2, or, in the formula:
Is a tetraamine as defined above for compounds of general formula (I), n'representing an integer of 3).

According to another embodiment, the invention also relates to a method of preparing a compound of general formula (Ik) as defined above, said method comprising general formula (V):
(In the formula,
And R ⁴ are as defined for the compound of general formula (I) above, and n′ represents an integer of 2, 3 and 4),
In the first step, it is reacted with an acid (eg, hydrochloric acid aqueous solution), and in the second step, a gadolinium(III) compound (eg, gadolinium oxide, gadolinium triacetate or gadolinium triacetate hydrate, gadolinium trichloride) Or gadolinium trinitrate) or its salt,
Thereby, the general formula (I−k):
(In the formula,
And R ⁴ are as defined for compounds of general formula (I) above, and n′ represents an integer of 2, 3 and 4).

Experimental section abbreviation

Materials and Instruments The chemicals used in the synthetic studies were of reagent grade quality and were used as received.

All reagents whose synthesis is not described in the experimental part are either commercially available or known compounds or may be produced from known compounds by methods known to those skilled in the art.

¹ H-NMR spectra were measured in CDCl ₃ , D ₂ O or DMSO-d ₆ respectively (room temperature, Bruker Avance 400 spectrometer, resonance frequency: 400.20 MHz (for ¹ H) or Bruker Avance 300 spectrometer, Resonance frequency: 300.13 MHz (for ¹ H) Chemical shifts are in ppm relative to an external standard (δ=0 ppm) sodium (trimethylsilyl)propionate-d ₄ (D ₂ O) or tetramethylsilane (DMSO-d ₆ ). It has been described in.

The compounds and intermediates produced according to the methods of the invention may require purification. Purification of organic compounds is well known to those skilled in the art, and there may be several ways to purify the same compound. In some cases, purification may not be necessary. In some cases, the compound may be purified by crystallization. In some cases, impurities may be removed by stirring with a suitable solvent. In some cases, the compound is, for example, a Biotage SNAP cartridge KP-Sil® or KP-NH in combination with a prepacked silica gel cartridge (eg, Biotage automated purification system (SP4® or Isolera Four®)). ®) and eluent (such as hexane/ethyl acetate or DCM/methanol gradient), in particular flash column chromatography. In some cases, the compound is loaded onto a suitable pre-packed reverse phase column and a trifluoroacetic acid, formic acid or aqueous ammonia solution, for example, using a Waters autopurifier equipped with a diode array detector and/or an online electrospray ionization mass spectrometer. May be purified by preparative HPLC in combination with an eluent such as a gradient of water and acetonitrile which may include additives such as.

The examples were analyzed and characterized by the following HPLC-based analytical methods measuring characteristic retention times and mass spectra.

Method 1: UPLC (ACN-HCOOH):
Measuring device: Waters Acquity UPLC-MS SQD 3001; Column: Acquity UPLC BEH C18 1.7 μm, 50×2.1 mm; Eluent A: water+0.1% formic acid, eluent B: acetonitrile; Gradient: 0-1. 6 min 1-99% B, 1.6-2.0 min 99% B; flow rate 0.8 mL/min; temperature: 60° C.; injection: 2 μl; DAD scan: 210-400 nm; ELSD.

Method 2: UPLC (ACN-HCOOH polarity):
Measuring device: Waters Acquity UPLC-MS SQD 3001; Column: Acquity UPLC BEH C18 1.7 μm, 50×2.1 mm; Eluent A: water+0.1% formic acid, eluent B: acetonitrile; Gradient: 0-1. 7 min 1-45% B, 1.7-2.0 min 45-99% B; flow rate 0.8 mL/min; temperature: 60° C.; injection: 2 μl; DAD scan: 210-400 nm; ELSD.

Method 3: UPLC (ACN-HCOOH Long Run):
Measuring instrument: Waters Acquity UPLC-MS SQD 3001; Column: Acquity UPLC BEH C18 1.7 μm, 50×2.1 mm; Eluent A: water+0.1% formic acid, eluent B: acetonitrile; Gradient: 0-4. 5 min 0-10% B; flow rate 0.8 mL/min; temperature: 60° C.; injection: 2 μl; DAD scan: 210-400 nm; ELSD.

How _{4: UPLC (ACN-NH 3} ):
Measuring instrument: Waters Acquity UPLC-MS ZQ2000; Column: Acquity UPLC BEH C18 1.7 μm, 50×2.1 mm; Eluent A: water+0.2% ammonia, eluent B: acetonitrile; Gradient: 0 to 1.6 min 1-99% B, 1.6-2.0 min 99% B; flow rate 0.8 mL/min; temperature: 60° C.; injection: 1 μL; DAD scan: 210-400 nm; ELSD.

Method 5: LC-MS:
Measuring instrument: Agilent 1290 UHPLCMS Tof; Column: BEH C 18 (Waters) 1.7 μm, 50×2.1 mm; Eluent A: Water+0.05 volume% formic acid (99%), Eluent B: Acetonitrile+0.05. % Formic acid; Gradient: 0-1.7min 98-10%A, 1.7-2.0min 10%A, 2.0-2.5min 10-98%A, Flow rate 1.2mL/min; Temperature: 60 C; DAD scan: 210-400 nm.

Example Compound Example 1
[4,10-bis(carboxylatomethyl)-7-{3,6,10,18,22,25-hexaoxo-26-[4,7,10-tris(carboxylatomethyl)-1,4,7 , 10-Tetraazacyclododecan-1-yl]-14-[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl] ] Propanoyl}amino)acetyl]-9,19-bis({[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl ] Propanoyl}amino)acetyl]amino}methyl)-4,7,11,14,17,21,24-heptaazaheptacosan-2-yl}-1,4,7,10-tetraazacyclododecane-1 -Ill] Acetate pentagadolinium

Example 1a
Di-tert-butyl(2-{[(2,5-dioxopyrrolidin-1-yl)oxy]carbonyl}propane-1,3-diyl)biscarbamate
3.60 g (11.3 mmol, 1 equivalent) of 3-[(tert-butoxycarbonyl)amino]-2-{[(tert-butoxycarbonyl)amino]methyl}propanoic acid (WO 2006/136460 (A2) No.) and 1.43 g (12.4 mmol, 1.1 eq) of 1-hydroxypyrrolidine-2,5-dione were dissolved in 120 mL of THF. To the reaction mixture was added dropwise a solution of 2.57 g (12.4 mmol, 1.1 eq) N,N'-dicyclohexylcarbodiimide in 60 mL THF. After stirring for 3 hours at room temperature, the resulting suspension was cooled to 0° C. and the precipitated urea was filtered off. The clear solution was evaporated to dryness to give 5.50 g (13.24 mmol, 117%) of the title compound.
UPLC (ACN-HCOOH): Rt. = 1.15 min.
MS (ES ⁺ ): m/z = 416.3 (M + H) ⁺ .

Example 1b
tert-Butyl (7,17-bis{[(tert-butoxycarbonyl)amino]methyl}-2,2-dimethyl-4,8,16-trioxo-3-oxa-5,9,12,15-tetraaza Octadecan-18-yl)carbamate
4.70 g (11.3 mmol, 2.22 equivalents) of di-tert-butyl(2-{[(2,5-dioxopyrrolidin-1-yl)oxy]carbonyl}propane-1,3-diyl)bis Carbamate (Example 1a) was dissolved in 120 mL of THF. To the reaction mixture, 0.53 g (5.10 mmol, 1 eq) N-(2-aminoethyl)ethane-1,2-diamine and 1.14 g (11.3 mmol, 2.22 eq) in 40 mL THF. Of triethylamine solution was added dropwise. After stirring for 3 hours at room temperature, the resulting suspension was diluted with dichloromethane. The organic solution was washed with aqueous sodium hydroxide solution (0.1M), water and dried over sodium sulfate. The crude product was isolated by evaporation under reduced pressure and purified by silica gel chromatography to give 2.81 g (3.99 mmol, 78%) of the title compound.
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ=1.36 (s, 36 H), 2.39-2.47 (m, 3 H), 2.52-2.58 (m, 4 H), 2.95-3.20 (m, 12 H), 6.64 (t, 4 H), 7.72 (t, 2 H) ppm.
UPLC (ACN-HCOOH): Rt. = 1.06 min.
MS (ES ⁺ ): m/z = 704.6 (M ⁺ +H).

Example 1c
N,N'-(iminodiethane-2,1-diyl)bis[3-amino-2-(aminomethyl)propanamide]pentahydrochloride
600 mg (0.85 mmol) tert-butyl (7,17-bis{[(tert-butoxycarbonyl)amino]methyl}-2,2-dimethyl-4,8,16-trioxo-3-oxa-5,9 , 12,15-Tetraazaoctadecan-18-yl)carbamate (Example 1b) was dissolved in 9.6 mL of methanol and 2.85 mL of aqueous hydrochloric acid solution (37%). The reaction mixture was heated at 50° C. for 2 hours under stirring. The suspension was evaporated to dryness for isolation to give 423 mg (0.87 mmol, 102%) of the title compound.
¹ H-NMR (400 MHz, D ₂ O): δ=3.04-3.15 (m, 2 H), 3.17-3.27 (m, 8 H), 3.29-3.38. (M, 4 H), 3.55 (t, 4 H) ppm.
UPLC (ACN-HCOOH): Rt. = 0.19 min.
MS (ES ⁺ ): m/z=304.2 (M+H) ⁺ , free base.

Example 1
[4,10-bis(carboxylatomethyl)-7-{3,6,10,18,22,25-hexaoxo-26-[4,7,10-tris(carboxylatomethyl)-1,4,7 , 10-Tetraazacyclododecan-1-yl]-14-[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl] ] Propanoyl}amino)acetyl]-9,19-bis({[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl ] Propanoyl}amino)acetyl]amino}methyl)-4,7,11,14,17,21,24-heptaazaheptacosan-2-yl}-1,4,7,10-tetraazacyclododecane-1 -Ill] Acetate pentagadolinium
150 mg (309 μmol, 1 eq) of N,N′-(iminodiethan-2,1-diyl)bis[3-amino-2-(aminomethyl)propanamide]pentahydrochloride (Example 1c) in 60 mL DMSO. Dissolved. 499 mg (3.86 mmol, 12.5 eq) N,N-diisopropylethylamine and 4.06 g (5.40 mmol, 17.5 eq) 2,2',2''-[10-(1-{[ 2-(4-Nitrophenoxy)-2-oxoethyl]amino}-1-oxopropan-2-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl]triacetate After adding gadolinium (see WO2001051095(A2)), the resulting reaction mixture was stirred and heated at 50° C. for 8 hours. The cooled solution was finally concentrated under reduced pressure to a volume of 15-20 mL. The concentrate was poured into 400 mL of ethyl acetate under stirring, the precipitate formed was filtered off and dried in vacuo. The solid was dissolved in water, the resulting solution was ultrafiltered with water using a 1 kDa membrane and the final concentrated water was lyophilized. The crude product was purified by RP chromatography to give 668 mg (64%, 199 μmol) of the title compound.
UPLC (ACN-HCOOH): Rt. = 0.46 min.
MS (ES ⁻ ): m/z (z=2)=1680. 5 (M−2H) ²⁻ ; (ES ⁺ ): m/z (z=3)=1121.3 (M+H) ³⁺ , m/ z (z=4)=841.4 [(M+H) ⁴⁺ .

Example 2
[4,10-bis(carboxylatomethyl)-7-{3,6,10,15,19,22-hexaoxo-23-[4,7,10-tris(carboxylatomethyl)-1,4,7 ,10-Tetraazacyclododecan-1-yl]-9,16-bis({[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclo Dodecane-1-yl]propanoyl}amino)acetyl]amino}methyl)-11-(2-{[3-{[({2-[4,7,10-tris(carboxylatomethyl)-1,4, 7,10-Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}-2-({[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7 , 10-Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)propanoyl]amino}ethyl)-4,7,11,14,18,21-hexaazatetracosan-2-yl} -1,4,7,10-Tetraazacyclododecan-1-yl]acetate hexagadolinium

Example 2a
tert-Butyl (12-{2-[(3-[(tert-butoxycarbonyl)amino]-2-{[(tert-butoxycarbonyl)amino]methyl}propanoyl)amino]ethyl}-7,14-bis{ [(Tert-Butoxycarbonyl)amino]methyl}-2,2-dimethyl-4,8,13-trioxo-3-oxa-5,9,12-triazapentadecan-15-yl)carbamate
890 mg (2.80 mmol, 3 equivalents) of 3-[(tert-butoxycarbonyl)amino]-2-{[(tert-butoxycarbonyl)amino]methyl}propanoic acid (see WO 2006/136460 (A2)). ) Was dissolved in 22 mL DMF. To the solution was added 434 mg (3.36 mmol, 3.6 eq) N,N-diisopropylethylamine and 1.28 g (3.36 mmol, 3.6 eq) HATU. The resulting reaction mixture was stirred at room temperature for 2 hours. 96.1 mg (0.93 mmol, 1 eq) N-(2-aminoethyl)ethane-1,2-diamine and 434 mg (3.36 mmol, 3.6 eq) N,N-diisopropyl in 9 mL DMF. After the solution of ethylamine was added dropwise, the resulting reaction mixture was heated under stirring at 70° C. for 3 hours. After cooling and diluting with dichloromethane, the solution was washed with aqueous sodium hydroxide solution (0.1M), aqueous citric acid solution (1%) and water, and dried over sodium sulfate. The crude product was isolated by evaporation under reduced pressure and purified by silica gel chromatography to give 451 mg (0.45 mmol, 48%) of the title compound.
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ=1.37 (s, 54 H), 2.36-2.49 (m, 3 H), 2.81-3.30 (m, 17 H), 3.36-3.70 (m, 3 H), 6.16-6.92 (m, 6 H), 7.77-8.35 (m, 2 H) ppm.
UPLC (ACN-HCOOH): Rt. = 1.49 min.
MS (ES ⁺ ): m/z = 1004.6 (M + H) ⁺ .

Example 2b
3-Amino-N,N-bis(2-{[3-amino-2-(aminomethyl)propanoyl]amino}ethyl)-2-(aminomethyl)propanamide hexahydrochloride
581 mg (0.58 mmol) tert-butyl (12-{2-[(3-[(tert-butoxycarbonyl)amino]-2-{[(tert-butoxycarbonyl)amino]methyl}propanoyl)amino]ethyl} -7,14-Bis{[(tert-butoxycarbonyl)amino]methyl}-2,2-dimethyl-4,8,13-trioxo-3-oxa-5,9,12-triazapentadecane-15-yl ) Carbamate (Example 2a) was dissolved in 9.3 mL of methanol and 2.9 mL of aqueous hydrochloric acid solution (37%). The reaction mixture was heated at 50° C. for 2 hours under stirring. The suspension was evaporated to dryness for isolation to give 376 mg (0.60 mmol, 103%) of the title compound.
¹ H-NMR (400 MHz, D ₂ O): δ=3.13-3.27 (m, 2 H), 3.28-3.85 (m, 21 H) ppm.
UPLC (ACN-HCOOH): Rt. = 0.19 min.
MS (ES ⁺ ): m/z = 404.3 (M+H) ⁺ , free base.

Example 2
[4,10-bis(carboxylatomethyl)-7-{3,6,10,15,19,22-hexaoxo-23-[4,7,10-tris(carboxylatomethyl)-1,4,7 ,10-Tetraazacyclododecan-1-yl]-9,16-bis({[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclo Dodecane-1-yl]propanoyl}amino)acetyl]amino}methyl)-11-(2-{[3-{[({2-[4,7,10-tris(carboxylatomethyl)-1,4, 7,10-Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}-2-({[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7 , 10-Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)propanoyl]amino}ethyl)-4,7,11,14,18,21-hexaazatetracosan-2-yl} -1,4,7,10-Tetraazacyclododecan-1-yl]acetate hexagadolinium
150 mg (241 μmol, 1 equivalent) of 3-amino-N,N-bis(2-{[3-amino-2-(aminomethyl)propanoyl]amino}ethyl)-2-(aminomethyl)propanamide hexahydrochloride (Example 2b) was dissolved in 60 mL DMSO. 467 mg (3.62 mmol, 15 eq) N,N-diisopropylethylamine and 3.80 g (5.06 mmol, 21 eq) 2,2',2''-[10-(1-{[2-(4 -Nitrophenoxy)-2-oxoethyl]amino}-1-oxopropan-2-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl]triacetato gadolinium (International Publication No. 2001051095 (see A2)) was added and the resulting reaction mixture was stirred and heated at 50° C. for 8 hours. The cooled solution was finally concentrated under reduced pressure to a volume of 15-20 mL. The concentrate was poured into 400 mL of ethyl acetate under stirring, the precipitate formed was filtered off and dried in vacuo. The solid was dissolved in water, the resulting solution was ultrafiltered with water using a 1 kDa membrane and the final concentrated water was lyophilized. The crude product was purified by RP chromatography to give 677 mg (166 μmol, 69%) of the title compound.
UPLC (ACN-HCOOH): Rt. = 0.44 min.
MS(ES ⁺ ): m/z(z=3)=11357.4(M+3H) ³⁺ , m/z(z=4)=1018.8(M+4H) ⁴⁺ ], m/z(z=5)= 815.7 (M+5H) ⁵⁺ .

Example 3
[4,10-bis(carboxylatomethyl)-7-{3,6,12,15-tetraoxo-16-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetra Azacyclododecan-1-yl]-9,9-bis({[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane-1- Ill]propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetraazaheptadecane-2-yl}-1,4,7,10-tetraazacyclododecan-1-yl]acetate tetra gadolinium
225 mg (1.65 mmol, 1 eq) of 2,2-bis(aminomethyl)propane-1,3-diamine (see W. Hayes et al., Tetrahedron 59 (2003), 7983-7996) was dissolved in 240 mL DMSO. . 1.71 g (13.2 mmol, 8 eq) N,N-diisopropylethylamine and 14.9 g (19.85 mmol, 12 eq) 2,2',2''-[10-(1-{[2- (4-Nitrophenoxy)-2-oxoethyl]amino}-1-oxopropan-2-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl]triacetato gadolinium ( After adding WO2001051095 (A2)), the resulting reaction mixture was stirred and heated at 50° C. for 8 hours. The cooled solution was finally concentrated under reduced pressure to a volume of 40-50 mL. The concentrate was poured into 600 mL of ethyl acetate under stirring, the precipitate formed was filtered off and dried in vacuo. The solid was dissolved in water, the resulting solution was ultrafiltered with water using a 1 kDa membrane and the final concentrated water was lyophilized. The crude product was purified by RP chromatography to give 3.42 g (80%, 1.33 mmol) of the title compound.
UPLC (ACN-HCOOH): Rt. = 0.42 min.
MS (ES ⁺ ): m/z (z = 2) = 12904 (M + H) 2 ⁺ , m / z (z = 3) = 860.7 (M + H) ^{3 +} .

Example 3 contains a mixture of stereoisomers, which stereoisomers exhibit the following absolute configurations:
All R, all S, RRRS, SSSR, RRSS.

Example 3-1
{4,10-bis(carboxylatomethyl)-7-[(2R,16R)-3,6,12,15-tetraoxo-16-[4,7,10-tris(carboxylatomethyl)-1,4 ,7,10-Tetraazacyclododecan-1-yl]-9,9-bis({[({(2R)-2-[4,7,10-tris(carboxylatomethyl)-1,4,7 , 10-Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetraazaheptadecane-2-yl]-1,4,7,10-tetra Azacyclododecan-1-yl}acetate tetragadolinium

Example 3-1a
tert-butyl {10,10-bis[({[(tert-butoxycarbonyl)amino]acetyl}amino)methyl]-2,2-dimethyl-4,7,13-trioxo-3-oxa-5,8, 12-triazatetradecane-14-yl}carbamate
See 2,2-bis(aminomethyl)propane-1,3-diamine tetrahydrochloride (851 mg, 3.06 mmol, 1 eq; in dichloromethane (50 mL); W. Hayes et al., Tetrahedron 59 (2003), 7983-7996. ) Of N,N-diisopropylethylamine (6.00 eq, 3.20 mL, 18.4 mmol) and 2,5-dioxopyrrolidin-1-yl N-(tert-butoxycarbonyl)glycinate (CAS No. [ 3392-07-2]; 6.00 eq, 5.00 g, 18.4 mmol) and stirred at room temperature for 2.5 days. The reaction mixture was diluted with water and the precipitate formed was filtered off and washed with water and dichloromethane. The precipitated material was chromatographed on silica gel (dichloromethane/methanol) to give the title compound (800 mg, 34%).
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ=1.36 (s, br, 36H), 2.74-2.76 (m, 8H), 3.48-3.50 (m, 8H), 6.96 (s, br, 0.4H*), 7.40-7.42 (m, 3.6H*), 7.91-8.00 (m, 4H) ppm.
LC-MS (ES ⁺ ): m/z=761.4 (M+H) ⁺ ; Rt. = 1.16 min.

Example 3-1b
2-amino-N-(3-[(aminoacetyl)amino]-2,2-bis{[(aminoacetyl)amino]methyl}propyl)acetamide tetrahydrochloride
Tert-Butyl {10,10-bis[({[(tert-butoxycarbonyl)amino]acetyl}amino)methyl]-2,2-dimethyl-4,7,13 from Example 11a in CPME (10 mL). A suspension of -trioxo-3-oxa-5,8,12-triazatetradecane-14-yl}carbamate (1.00 eq, 800 mg, 1.05 mmol) was cooled to 0 °C and CPME (10 eq, Treated drop-wise with HCl in 3.5 mL of 3M solution, 10.5 mmol). The reaction mixture was stirred at 0 °C for 1 h and at room temperature overnight, to which was added dioxane (4 mL) and another amount of HCl in CPME (30 eq, 11 mL of 3M solution, 32 mmol) and stirred at room temperature for 2 days. Continued. The resulting suspension was concentrated in vacuo to give the title compound (575mg, quantitative). This compound was not further purified.
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ=3.17-3.18 (m, 8H), 3.59-3.61 (m, 8H), 8.21 (s, br, 12H), 8.55 (t, 4H) ppm.
LC-MS (ES ⁺ ): m/z=361.2 (M-3HCl-Cl ^- ) ⁺ ; Rt. = 0.10 min.

Example 3-1c
Benzyl(2S)-2-{[(trifluoromethyl)sulfonyl]oxy}propanoate
H. C. J. Prepared according to Ottenheim et al., Tetrahedron 44 (1988), 5583-5595: (S)-(−)-lactic acid benzyl ester (CAS No. [56777-24-3]; 1.00) in dry dichloromethane (95 mL). Equivalent, 5.00 g, 27.7 mmol) was cooled to 0° C. and trifluoromethanesulfonic anhydride (CAS No. [358-23-6]; 1.1 eq, 5.2 mL, 8.6 g, 31 mmol). After stirring for 5 minutes, 2,6-dimethylpyridine (1.15 equivalents, 3.72 mL, 3.42 g) was added and stirring was continued for another 5 minutes. The resulting reaction mixture was used directly in the next step.

Example 3-1d
Benzyl(2R)-2-[4,7,10-tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]propanoate
Tri-tert-butyl 2,2′,2″-(1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate (CAS No. [122555] in dry dichloromethane (75 mL). -91-3]; 1.00 equiv., 9.52 g, 18.5 mmol) was cooled to 0°C and benzyl(2S)-2-{[(tris in dichloromethane prepared in Example 3-1c. Fluoromethyl)sulfonyl]oxy}propanoate; and N,N-diisopropylethylamine (3.0 eq, 9.7 mL, 55 mmol). The resulting solution was stirred at room temperature for 6 days, diluted with ethyl acetate and washed with saturated aqueous sodium hydrogen carbonate solution. The organic layer was dried over sodium sulfate and concentrated under reduced pressure. The resulting material was purified by amino phase silica gel chromatography (KP-NH®, hexane/ethyl acetate to dichloromethane/methanol) to give the title compound (1.92 g, 14%).
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ=1.20 (d, 3H), 1.37-1.45 (m, 27H), 1.98-2.01 (m, 3H) , 2.08-2.24 (m, 5H), 2.57-2.84 (m, 7H), 2.94-3.11 (m, 4H), 3.38-3.48 (m, 3H), 3.75 (q, 1H), 5.07-5.17 (m, 2H), 7.32-7.40 (m, 5H) ppm.
LC-MS (ES ⁺ ): m/z=677.5 (M+H) ⁺ , m/z (z=2)=339.2 (M+H) ²⁺ ; Rt. = 1.06 min.

Example 3-1e
(2R)-2-[4,7,10-Tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]propanoic acid
Benzyl(2R)-2-[4,7,10-tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1- in methanol (17.5 mL) A solution of il]propanoate (Example 3-1d; 1.92 g, 2.84 mmol) was treated with Pd/C (10 wt%; 0.050 eq, 151 mg, 0.14 mmol) at room temperature under hydrogen atmosphere. Stir for 20 hours. The reaction mixture was filtered over Celite®, washed with methanol and the filtrate concentrated in vacuo to give the title compound (1.51 g, 88%). This compound was not further purified.
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ=1.11 (s, br, 3H), 1.42-1.43 (m, 27H), 1.97-2.13 (m, 5H), 2.56-2.82 (m, 7H), 2.97-3.07 (m, 4H), 3.34-3.53 (m, 7H), 12.8 (s, br, 1H) ppm.
_{UPLC (ACN-NH 3):} Rt. = 1.31 min.
MS (ES ⁺ ): m/z=587 (M+H) ⁺ .
LC-MS (ES ⁺ ): m/z=587 (M+H) ⁺ , m/z (z=2)=294.2 (M+H) ²⁺ ; Rt. = 0.79 min.

Example 3-1f
tert-butyl {4,10-bis(2-tert-butoxy-2-oxoethyl)-7-[(2R,16R)-3,6,12,15-tetraoxo-16-[4,7,10-tris (2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]-9,9-bis({[({(2R)-2-[4,7 , 10-Tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)-4,7,11, 14-Tetraazaheptadecane-2-yl]-1,4,7,10-tetraazacyclododecan-1-yl}acetate
(2R)-2-[4,7,10-Tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecane- in N,N-dimethylacetamide (15 mL) 1-yl]propanoic acid (Example 3-1e; 12.0 eq, 1.50 g, 2.56 mmol) was treated with HATU (14.4 eq, 1.17 g, 3.07 mmol) and N,N-diisopropyl. Treated with ethylamine (14.4 eq, 534 μL, 3.07 mmol) and stirred at room temperature for 20 minutes. 2-Amino-N-(3-[(aminoacetyl)amino]-2,2-bis{[(aminoacetyl)amino]methyl}propyl)acetamide tetrahydrochloride in N,N-dimethylacetamide (6 mL) ( Example 3-1b; 1.00 eq, 108 mg, 213 μmol) suspension was added and the resulting mixture was stirred at 50° C. overnight. The reaction mixture was concentrated under reduced pressure and the obtained residue was subjected to amino phase silica gel chromatography (KP-NH (registered trademark), ethyl acetate to ethyl acetate/methanol) to obtain the title compound (260 mg, 42%). ..
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ=1.03 (s, br, 5H), 1.28 (s, br, 7H), 1.36-1.43 (m, 108H) , 1.87-2.24 (m, 23H), 2.42 (s, br, 4H), 2.53-2.84 (m, 41H), 2.97-3.18 (m, 17H) , 3.28 (s, br, 5H), 3.39-3.46 (m, 6H), 3.58 (s, br, 7H), 3.76 (s, br, 2H), 4.01. (S, br, 3H), 7.81 (s, br, 5H), 8.33 (s, br, 2H), 9.27 (s, br, 1H) ppm.
_{UPLC (ACN-NH 3):} Rt. = 1.23 min.
MS (ES ⁺ ): m/z (z = 4) = 660 (M + H) ^{4 +} .
LC-MS (ES ⁺ ): m/z (z=2)=1318 (M+H) ²⁺ , m/z (z=3)=879 (M+H) ³⁺ , m/z (z=4)=660 (M+H) ) ⁴⁺ ; Rt. = 0.94 min.

Example 3-1g
{4,10-bis(carboxymethyl)-7-[(2R,16R)-3,6,12,15-tetraoxo-16-[4,7,10-tris(carboxymethyl)-1,4,7 , 10-Tetraazacyclododecan-1-yl]-9,9-bis({[({(2R)-2-[4,7,10-tris(carboxymethyl)-1,4,7,10- Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetraazaheptadecane-2-yl]-1,4,7,10-tetraazacyclododecane -1-yl}acetic acid
tert-butyl {4,10-bis(2-tert-butoxy-2-oxoethyl)-7-[(2R,16R)-3,6,12,15-tetraoxo-16-[4,7,10-tris (2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]-9,9-bis({[({(2R)-2-[4,7 , 10-Tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)-4,7,11, 14-Tetraazaheptadecane-2-yl]-1,4,7,10-tetraazacyclododecan-1-yl}acetate (Example 3-1f; 260 mg, 0.099 mmol) was stirred under stirring with TFA (25 mL). ) At room temperature overnight. The reaction mixture was concentrated under reduced pressure and the resulting residue was taken up in water (20 mL) and freeze dried. The crude product was used in the next chemical step without further characterization.

Example 3-1
{4,10-bis(carboxylatomethyl)-7-[(2R,16R)-3,6,12,15-tetraoxo-16-[4,7,10-tris(carboxylatomethyl)-1,4 ,7,10-Tetraazacyclododecan-1-yl]-9,9-bis({[({(2R)-2-[4,7,10-tris(carboxylatomethyl)-1,4,7 , 10-Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetraazaheptadecane-2-yl]-1,4,7,10-tetra Azacyclododecan-1-yl}acetate tetragadolinium Crude product from Example 3-1g {4,10-bis(carboxymethyl)-7-[(2R,16R)-3,6,12,15-tetraoxo -16-[4,7,10-Tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl]-9,9-bis({[({(2R)-2- [4,7,10-Tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetra Azaheptadecane-2-yl]-1,4,7,10-tetraazacyclododecan-1-yl}acetic acid was dissolved in water (20 mL). Tris(acetato-κO) gadolinium tetrahydrate (298 mg, 0.734 mmol) was added and the reaction mixture was stirred at 70° C. for 2 hours. The pH value of the resulting solution was adjusted to 4.5 by adding aqueous sodium hydroxide solution (2N), and stirring was continued at 70°C for 2 days. The resulting solution was ultrafiltered through water (7×100 mL) using a 1 kDa membrane and the final concentrated water was lyophilized to give the title compound (70 mg, 27% over 2 steps).
UPLC (ACN-HCOOH): Rt. = 0.39 min.
MS (ES ⁺ ): m/z (z = 2) = 129.1 (M + H) 2 ⁺ , m / z (z = 3) = 860.3 (M + H) ^{3 +} .
LC-MS (ES ⁺ ): m/z (z = 2) = 129.3 (M + H) 2 ⁺ , m / z (z = 3) = 860.9 (M + H) ^{3 +} , m / z (z = 4) = 645.6 (M+H) ⁴⁺ ; Rt. = 0.25 min.

Example 3-2
{4,10-bis(carboxylatomethyl)-7-[(2S,16S)-3,6,12,15-tetraoxo-16-[4,7,10-tris(carboxylatomethyl)-1,4 ,7,10-Tetraazacyclododecan-1-yl]-9,9-bis({[({(2S)-2-[4,7,10-tris(carboxylatomethyl)-1,4,7 , 10-Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetraazaheptadecane-2-yl]-1,4,7,10-tetra Azacyclododecan-1-yl}acetate tetragadolinium

Example 3-2a
Benzyl (2R)-2-{[(trifluoromethyl)sulfonyl]oxy}propanoate
Similar to the corresponding S-isomer (Example 3-1c), (R)-(+)-lactic acid benzyl ester (CAS No. [74094-05-6]; 8.00 g, 44.4 mmol) in dichloromethane. ) Was prepared. The resulting reaction mixture was used directly in the next step.

Example 3-2b
Benzyl(2S)-2-[4,7,10-tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]propanoate
Similar to the corresponding R-isomer (Example 3-1d), tri-tert-butyl 2,2′,2″-(1,4,7,10-tetraazacyclododecane-1,4,7 -Triyl)triacetate (CAS No. [122555-91-3]; 1.00 eq, 15.2 g, 29.6 mmol) and benzyl(2R)-2-{[ in dichloromethane prepared in Example 3-2a. Prepared from the reaction mixture of (trifluoromethyl)sulfonyl]oxy}propanoate.
LC-MS (ES ⁺ ): m/z=677.4 (M+H) ⁺ , m/z (z=2)=339.2 (M+H) ²⁺ ; Rt. = 0.94 min.

Example 3-2c
(2S)-2-[4,7,10-Tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]propanoic acid
Similar to the corresponding R-isomer (Example 3-1e), benzyl(2S)-2-[4,7,10-tris(2-tert-butoxy-2-oxoethyl)-1,4,7, Prepared from 10-tetraazacyclododecan-1-yl]propanoate (Example 3-2b).
_{UPLC (ACN-NH 3):} Rt. = 1.31 min.
MS (ES ⁺ ): m/z=587 (M+H) ⁺ .
LC-MS (ES ⁺ ): m/z=587.4 (M+H) ⁺ , m/z (z=2)=294.2 (M+H) ²⁺ ; Rt. = 0.82 min.

Example 3-2d
tert-Butyl {4,10-bis(2-tert-butoxy-2-oxoethyl)-7-[(2S,16S)-3,6,12,15-tetraoxo-16-[4,7,10-tris (2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]-9,9-bis({[({(2S)-2-[4,7 , 10-Tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)-4,7,11, 14-Tetraazaheptadecane-2-yl]-1,4,7,10-tetraazacyclododecan-1-yl}acetate
Similar to the corresponding R-isomer (Example 3-1f), (2S)-2-[4,7,10-tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10 -Tetraazacyclododecan-1-yl]propanoic acid (Example 3-2c) and 2-amino-N-(3-[(aminoacetyl)amino]-2,2-bis{[(aminoacetyl)amino] Prepared from methyl}propyl)acetamide tetrahydrochloride (Example 3-1b).
LC-MS (ES ⁺ ): m/z (z=2)=1318 (M+H) ²⁺ , m/z (z=3)=879 (M+H) ³⁺ , m/z (z=4)=660 (M+H) ) ⁴⁺ ; Rt. = 0.95 min.

Example 3-2e
{4,10-bis(carboxymethyl)-7-[(2S,16S)-3,6,12,15-tetraoxo-16-[4,7,10-tris(carboxymethyl)-1,4,7 , 10-Tetraazacyclododecan-1-yl]-9,9-bis({[({(2S)-2-[4,7,10-tris(carboxymethyl)-1,4,7,10- Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetraazaheptadecane-2-yl]-1,4,7,10-tetraazacyclododecane -1-yl}acetic acid
Similar to the corresponding R-isomer (Example 3-1g), tert-butyl {4,10-bis(2-tert-butoxy-2-oxoethyl)-7-[(2S,16S)-3,6 ,12,15-Tetraoxo-16-[4,7,10-tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]-9,9 -Bis({[({(2S)-2-[4,7,10-tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl] Propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetraazaheptadecane-2-yl]-1,4,7,10-tetraazacyclododecan-1-yl}acetate (Example 3-2d). The crude product was used in the next chemical step without further characterization.

Example 3-2
{4,10-bis(carboxylatomethyl)-7-[(2S,16S)-3,6,12,15-tetraoxo-16-[4,7,10-tris(carboxylatomethyl)-1,4 ,7,10-Tetraazacyclododecan-1-yl]-9,9-bis({[({(2S)-2-[4,7,10-tris(carboxylatomethyl)-1,4,7 , 10-Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetraazaheptadecane-2-yl]-1,4,7,10-tetra Azacyclododecan-1-yl}acetate tetragadolinium Similar to the corresponding R-isomer (Example 3-1), {4,10-bis(carboxymethyl)-7-[(2S,16S)-3, 6,12,15-Tetraoxo-16-[4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl]-9,9-bis({[( {(2S)-2-[4,7,10-Tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)-4, 7,11,14-Tetraazaheptadecan-2-yl]-1,4,7,10-tetraazacyclododecan-1-yl}acetic acid (Example 3-2e) and tris(acetato-κO)gadolinium tetra Prepared from hydrate at pH 4.5. The resulting reaction solution was ultrafiltered with water (8 x 100 mL) using a 1 kDa membrane and the final concentrated water was lyophilized and purified by preparative HPLC.
UPLC (ACN-HCOOH): Rt. = 0.41 min.
MS (ES ⁺ ): m/z (z=2)=1290 (M+H) ²⁺ , m/z (z=3)=861 (M+H) ³⁺ .
LC-MS (ES ⁺ ): m/z (z=2)=1290 (M+H) ²⁺ , m/z (z=3)=860 (M+H) ³⁺ , m/z (z=4)=645.6 (M+H) ⁴⁺ ; Rt. = 0.23 min.

Example 4
[4-(1-{[2-(bis{2-[({1,4-bis[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7,10- Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]-1,4-diazepan-6-yl}carbonyl)amino]ethyl}amino)-2-oxoethyl]amino}-1-oxopropan-2-yl )-7,10-Bis(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetate pentagadolinium

Example 4a
6-(methoxycarbonyl)-1,4-diazepandiium dichloride
6.00 g (17.7 mmol) of methyl 1,4-dibenzyl-1,4-diazepan-6-carboxylate [see US Pat. No. 5,866,562] was dissolved in 30 mL of methanol. After addition of 6 mL aqueous hydrochloric acid (37%), 6 mL water and 600 mg palladium on charcoal (10%), the reaction mixture was hydrogenated (1 atm) at 40° C. for 17 hours. The catalyst was filtered off and the solution was evaporated under reduced pressure to give 4.1 g (17.7 mmol, 100%) of the title compound.
¹ H-NMR (400 MHz, D ₂ O): δ=3.62-3.84 (m, 9 H), 3.87 (s, 3 H) ppm.
UPLC (ACN-HCOOH): Rt. = 0.20 min.
MS (ES ⁺ ): m/z = 159.1 (M+H) ⁺ , free base.

Example 4b
1,4-di-tert-butyl 6-methyl 1,4-diazepane-1,4,6-tricarboxylate
4.00 g (17.3 mmol, 1 eq) of 6-(methoxycarbonyl)-1,4-diazepandiium dichloride (Example 4a) was dissolved in 80 mL of DMF. After adding 7.71 g (76.2 mmol, 4.4 eq) trimethylamine and 8.31 g (38.1 mmol, 2.2 eq) di-tert-butyl dicarbonate, the resulting reaction mixture was allowed to stand at room temperature. Stir overnight. The suspension was filtered, the filtrate evaporated under reduced pressure and diluted with ethyl acetate. The resulting solution was washed with aqueous citric acid solution (pH=3-4), semisaturated aqueous sodium bicarbonate solution, dried over sodium sulfate and evaporated under reduced pressure to give 4.92 g (13.7 mmol, 79%) of the title compound. Got
¹ H-NMR (300 MHz, DMSO-d ₆ ): δ=1.36 (s, 18 H), 2.69-3.27 (m, 4 H), 3.35-4.00 (m, 5 H), 3.62 (s, 3 H) ppm.
UPLC (ACN-HCOOH): Rt. = 1.32 min.
MS (ES ⁺ ): m/z = 359.2 (M + H) ⁺ .

Example 4c
1,4-bis(tert-butoxycarbonyl)-1,4-diazepan-6-carboxylic acid
4.86 g (13.66 mmol) of 1,4-di-tert-butyl 6-methyl 1,4-diazepane-1,4,6-tricarboxylate (Example 4b) were dissolved in 82 mL of THF. After addition of 27 mL aqueous sodium hydroxide solution (2M), the resulting reaction mixture was stirred at room temperature for 20 hours, diluted with water and citric acid was added to make it acidic (pH=3-4). The crude product was extracted with dichloromethane, the organic layer was washed with brine, dried over sodium sulfate and evaporated to dryness to give 4.67 g (12.4 mmol, 91%) of the title compound.
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ=1.38 (s, 18 H), 2.58-2.86 (m, 1 H), 2.94-4.00 (m, 8 H), 12.50 (s, br, 1 H) ppm.
UPLC (ACN-HCOOH): Rt. = 1.12 min.
MS (ES ⁺ ): m/z = 345.2 (M + H) ⁺ .

Example 4d
Di-tert-butyl 6-{[(2,5-dioxopyrrolidin-1-yl)oxy]carbonyl}-1,4-diazepan-1,4-dicarboxylate
1.76 g (5.11 mmol, 1 eq) of 1,4-bis(tert-butoxycarbonyl)-1,4-diazepan-6-carboxylic acid (Example 4c) and 0.65 g (5.62 mmol, 1. 1 equivalent of 1-hydroxypyrrolidine-2,5-dione was dissolved in 50 mL of THF. A solution of 1.16 g (5.62 mmol, 1.1 eq) N,N'-dicyclohexylcarbodiimide in 30 mL THF was added and the resulting reaction mixture was refluxed for 5 hours. The suspension was cooled to 0° C. and the precipitated urea was filtered off. The final solution of activated ester was used directly in the next chemical step.
UPLC (ACN-HCOOH): Rt. = 1.24 min.
MS (ES ⁺ ): m/z=442.3 (M+H) ⁺ .

Example 4e
Tetra-tert-butyl 6,6'-[iminobis(ethane-2,1-diylcarbamoyl)]bis(1,4-diazepane-1,4-dicarboxylate)
Activated ester from Example 4d (5.11 mmol, 2.2 eq) di-tert-butyl 6-{[(2,5-dioxopyrrolidin-1-yl)oxy]carbonyl}-1,4-diazepane In a solution of -1,4-dicarboxylate, 517 mg (5.11 mmol, 2.2 eq) triethylamine and 240 mg (2.32 mmol, 1 eq) N-(2-aminoethyl)ethane-1,2- Diamine was added. The resulting reaction mixture was stirred at room temperature for 20 hours and diluted with dichloromethane. The solution was washed with aqueous sodium hydroxide solution (0.1M), water and dried over sodium sulfate. The crude product was isolated by evaporation and purified by silica gel chromatography to give 1.20 g (1.59 mmol, 68%) of the title compound.
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ=1.37 (s, 36 H), 2.51-2.70 (m, 7 H), 2.85-3.28 (m, 12 H), 3.45-4.10 (m, 8 H), 7.69-8.27 (m, 2 H) ppm.
UPLC (ACN-HCOOH): Rt. = 1.20 min.
MS (ES ⁺ ): m/z=756.7 (M+H) ⁺ .

Example 4f
N,N'-(iminodiethane-2,1-diyl)bis(1,4-diazepan-6-carboxamide)pentahydrochloride
385 mg (0.51 mmol) of tetra-tert-butyl 6,6'-[iminobis(ethane-2,1-diylcarbamoyl)]bis(1,4-diazepane-1,4-dicarboxylate) (Example 4e) Was dissolved in 5.7 mL of methanol and 1.7 mL of aqueous hydrochloric acid solution (37%). The reaction mixture was heated at 50° C. for 2 hours under stirring. The suspension was evaporated to dryness for isolation to give 277 mg (0.51 mmol, 100%) of the title compound.
¹ H-NMR (400 MHz, D ₂ O): δ=3.18 (t, 4 H), 3.32-3.40 (m, 2 H), 3.51 (t, 4 H), 3 57-3.69 (m, 16 H) ppm.
UPLC (ACN-HCOOH): Rt. = 0.24 min.
MS (ES ⁺ ): m/z=356.3 (M+H) ⁺ , free base.

Example 4
[4-(1-{[2-(bis{2-[({1,4-bis[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7,10- Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]-1,4-diazepan-6-yl}carbonyl)amino]ethyl}amino)-2-oxoethyl]amino}-1-oxopropan-2-yl )-7,10-Bis(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetate pentagadolinium
150 mg (279 μmol, 1 eq) N,N′-(iminodiethane-2,1-diyl)bis(1,4-diazepan-6-carboxamide)pentahydrochloride (Example 4f) were dissolved in 60 mL DMSO. 451 mg (3.49 mmol, 12.5 eq) N,N-diisopropylethylamine and 3.67 g (4.88 mmol, 17.5 eq) 2,2',2''-[10-(1-{[ 2-(4-Nitrophenoxy)-2-oxoethyl]amino}-1-oxopropan-2-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl]triacetate After adding gadolinium (see WO2001051095(A2)), the resulting reaction mixture was stirred and heated at 50° C. for 8 hours. The cooled solution was finally concentrated under reduced pressure to a volume of 15-20 mL. The concentrate was poured into 400 mL of ethyl acetate under stirring, the precipitate formed was filtered off and dried in vacuo. The solid was dissolved in water, the resulting solution was ultrafiltered with water using a 1 kDa membrane and the final concentrated water was lyophilized. The crude product was purified by RP chromatography to give 672 mg (197 μmol, 70%) of the title compound.
UPLC (ACN-HCOOH): Rt. = 0.43 min.
MS (ES ⁻ ): m/z (z=2)=1706. 3 (M−2H) ²⁻ m; (ES ⁺ ): m/z (z=4)=854.5 (M+4H) ⁴⁺ .

Example 5
2, 2', 2'', 2''', 2'''', 2''''', 2'''''', 2''''''', 2'''''''',2''''''''',2'''''''''',2''''''''''',2'''''''''''',2''''''''''''',2'''''''''''''',2''''''''''''''',2'''''''''''''''',2'''''''''''''''''-{ethane-1,2-diylcarbamoyl-1 ,4-Diazepane-6,1,4-triyltris[(2-oxoethane-2,1-diyl)imino(1-oxopropane-1,2-diyl)-1,4,7,10-tetraazacyclododecane −10,1,4,7-Tetrayl]}octadecaacetate hexagadolinium

Example 5a
Hexa-tert-butyl 6,6′,6″-(ethane-1,2-diylcarbamoyl)tris(1,4-diazepan-1,4-dicarboxylate)
1.20 g (3.48 mmol, 3 eq) of 1,4-bis(tert-butoxycarbonyl)-1,4-diazepan-6-carboxylic acid (Example 4c), 540 mg (4.18 mmol, 3.6 eq) ) Diisopropylethylamine and 1.59 g (4.18 mmol, 3.6 eq) HATU were dissolved in 30 mL DMF and stirred at room temperature for 2 hours. Of 120 mg (1.16 mmol, 1 eq) N-(2-aminoethyl)ethane-1,2-diamine and 540 mg (4.18 mmol, 3.6 eq) N,N-diisopropylethylamine in 8 mL DMF. After the solution was added dropwise, the resulting reaction mixture was heated under stirring at 70° C. for 3 hours. After cooling and diluting with dichloromethane, the solution was washed with aqueous sodium hydroxide solution (0.1M), aqueous citric acid solution (1%), water and dried over sodium sulfate. The crude product was isolated by evaporation under reduced pressure and purified by silica gel chromatography to give 660 mg (0.61 mmol, 52%) of the title compound.
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ=1.38 (s, 54 H), 2.55-4.06 (m, 35 H), 7.90-8.52 (m, 2 H) ppm.
UPLC (ACN-HCOOH): Rt. = 1.64 min.
MS (ES ⁺ ): m/z=1082.7 (M+H) ⁺ .

Example 5b
N,N-bis{2-[(1,4-diazepan-6-ylcarbonyl)amino]ethyl}-1,4-diazepan-6-carboxamide hexahydrochloride
654 mg (0.60 mmol) hexa-tert-butyl 6,6′,6″-(ethane-1,2-diylcarbamoyl)tris(1,4-diazepan-1,4-dicarboxylate) (Example 5a) was dissolved in 6.8 mL methanol and 3 mL aqueous hydrochloric acid solution (37%). The reaction mixture was heated at 50° C. for 2.5 hours with stirring. The suspension was evaporated to dryness for isolation to give 441 mg (0.63 mmol, 105%) of the title compound.
¹ H-NMR (400 MHz, DMSO-d ₆ ): δ=3.20-3.71 (m, 35 H), 8.50-8.80 ppm (m, 2 H), 9.76 (s). , Br, 12 H).
UPLC (ACN-HCOOH): Rt. = 0.19 min.
MS (ES ⁺ ): m/z = 482.3 (M+H) ⁺ , free base.

Example 5
2, 2', 2'', 2''', 2'''', 2''''', 2'''''', 2''''''', 2'''''''',2''''''''',2'''''''''',2''''''''''',2'''''''''''',2''''''''''''',2'''''''''''''',2''''''''''''''',2'''''''''''''''',2'''''''''''''''''-{ethane-1,2-diylcarbamoyl-1 ,4-Diazepane-6,1,4-triyltris[(2-oxoethane-2,1-diyl)imino(1-oxopropane-1,2-diyl)-1,4,7,10-tetraazacyclododecane −10,1,4,7-Tetrayl]}octadecaacetate hexagadolinium
150 mg (214 μmol, 1 eq) of N,N-bis{2-[(1,4-diazepan-6-ylcarbonyl)amino]ethyl}-1,4-diazepan-6-carboxamide hexahydrochloride (Example 5b) ) Was dissolved in 60 mL DMSO. 0.42 g (3.21 mmol, 15 eq) of N,N-diisopropylethylamine and 3.38 g (4.50 mmol, 21 eq) of 2,2',2''-[10-(1-{[2- (4-Nitrophenoxy)-2-oxoethyl]amino}-1-oxopropan-2-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl]triacetato gadolinium ( After adding WO2001051095 (A2)), the resulting reaction mixture was stirred and heated at 50° C. for 8 hours. The cooled solution was finally concentrated under reduced pressure to a volume of 15-20 mL. The concentrate was poured into 400 mL of ethyl acetate under stirring, the precipitate formed was filtered off and dried in vacuo. The solid was dissolved in water, the resulting solution was ultrafiltered with water using a 1 kDa membrane and the final concentrated water was lyophilized. The crude product was purified by RP chromatography to give 595 mg (143 μmol, 67%) of the title compound.
UPLC (ACN-HCOOH): Rt. = 0.41 min.
MS(ES ⁺ ): m/z(z=3)=11384.6(M+H) ³⁺ , m/z(z=4)=1039.5(M+H) ⁴⁺ , m/z(z=5)=831 .6 (M+H) ⁵⁺ .

Example 6
2, 2', 2'', 2''', 2'''', 2''''', 2'''''', 2''''''', 2'''''''',2''''''''',2'''''''''',2''''''''''',2'''''''''''',2''''''''''''',2'''''''''''''',2''''''''''''''',2'''''''''''''''',2'''''''''''''''''-(1,4,7-triazonan-1,-1, 4,7-Triyltris{carbonyl-1,4-diazepan-6,1,4-triylbis[(2-oxoethane-2,1-diyl)imino(1-oxopropane-1,2-diyl)-1,4 , 7,10-Tetraazacyclododecane-10,1,4,7-tetrayl]}) octadecaacetate hexagadolinium

Example 6a
Hexa-tert-butyl 6,6′,6″-(1,4,7-triazonan-1,4,7-triyltricarbonyl)tris(1,4-diazepan-1,4-dicarboxylate)
800 mg (2.32 mmol, 3 eq) 1,4-bis(tert-butoxycarbonyl)-1,4-diazepan-6-carboxylic acid (Example 4c), 360 mg (2.79 mmol, 3.6 eq) Diisopropylethylamine and 1.06 g (2.79 mmol, 3.6 eq) HATU were dissolved in 20 mL DMF and stirred at room temperature for 2 hours. After a solution of 100 mg (774 μmol, 1 eq) 1,4,7-triazonan trihydrochloride and 360 mg (2.79 mmol, 3.6 eq) N,N-diisopropylethylamine in 5 mL DMF was added dropwise, The resulting reaction mixture was heated at 70° C. for 3 hours with stirring. After cooling and diluting with dichloromethane, the solution was washed with aqueous sodium hydroxide solution (0.1M), aqueous citric acid solution (1%), water and dried over sodium sulfate. The crude product was isolated by evaporation under reduced pressure and purified by silica gel chromatography to give 545 mg (492 μmol, 63%) of the title compound.
¹ H-NMR (400 MHz, CDCl ₃ ): δ=1.47 (s, 54 H), 2.85-4.45 (m, 39 H) ppm.
UPLC (ACN-HCOOH): Rt. = 1.73 min.
MS (ES ⁺ ): m/z = 1108.8 (M+H) ⁺ .

Example 6b
1,4,7-Triazonan-1,4,7-triyltris(1,4-diazepan-6-ylmethanone) hexahydrochloride
380 mg (343 μmol) hexa-tert-butyl 6,6′,6″-(1,4,7-triazonan-1,4,7-triyltricarbonyl)tris(1,4-diazepan-1,4-) The dicarboxylate) (Example 6a) was dissolved in 3.90 mL of methanol and 1.72 mL of aqueous hydrochloric acid solution (37%). The reaction mixture was heated at 50° C. for 2.5 hours with stirring. The suspension was evaporated to dryness for isolation to give 257 mg (354 μmol, 103%) of the title compound.
UPLC (ACN-HCOOH): Rt. = 0.19 min.
MS (ES ⁺ ): m/z=508.4 (M+H) ⁺ , free base.

Example 6
2, 2', 2'', 2''', 2'''', 2''''', 2'''''', 2''''''', 2'''''''',2''''''''',2'''''''''',2''''''''''',2'''''''''''',2''''''''''''',2'''''''''''''',2''''''''''''''',2'''''''''''''''',2'''''''''''''''''-(1,4,7-triazonan-1,-1, 4,7-Triyltris{carbonyl-1,4-diazepan-6,1,4-triylbis[(2-oxoethane-2,1-diyl)imino(1-oxopropane-1,2-diyl)-1,4 , 7,10-Tetraazacyclododecane-10,1,4,7-tetrayl]}) octadecaacetate hexagadolinium
175 mg (241 μmol, 1 eq) of 1,4,7-triazonan-1,4,7-triyltris(1,4-diazepan-6-ylmethanone) hexahydrochloride (Example 6b) was dissolved in 60 mL DMSO. 467 mg (3.61 mmol, 15 eq) N,N-diisopropylethylamine and 3.80 g (5.06 mmol, 21 eq) 2,2',2''-[10-(1-{[2-(4 -Nitrophenoxy)-2-oxoethyl]amino}-1-oxopropan-2-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl]triacetato gadolinium (International Publication No. 2001051095 (A2)) was added and the resulting reaction mixture was stirred and heated at 50° C. for 8 hours. The cooled solution was finally concentrated under reduced pressure to a volume of 15-20 mL. The concentrate was poured into 400 mL of ethyl acetate under stirring, the precipitate formed was filtered off and dried in vacuo. The solid was dissolved in water, the resulting solution was ultrafiltered with water using a 1 kDa membrane and the final concentrated water was lyophilized. The crude product was purified by RP chromatography to give 590 mg (141 μmol, 58%) of the title compound.
UPLC (ACN-HCOOH): Rt. = 0.43 min.
MS(ES ⁺ ): m/z(z=3)=1393.1(M+3H) ³⁺ , m/z(z=4)=1045.5(M+4H) ⁴⁺ , m/z(z=5)=837 .0 [(M+5H) ⁵⁺ .

Example 7
2, 2', 2'', 2''', 2'''', 2''''', 2'''''', 2''''''', 2'''''''',2''''''''',2'''''''''',2'''''''''''-{1,4,7,10-tetra Azacyclododecane-1,4,7,10-tetrayltetrakis[(2-oxoethane-2,1-diyl)imino(1-oxopropane-1,2-diyl)-1,4,7,10-tetra Azacyclododecane-10,1,4,7-tetrayl]}dodecaacetate tetragadolinium
35 mg (203 μmol, 1 eq) of 1,4,7,10-tetraazacyclododecane were dissolved in 60 mL DMSO. 2.14 g (2.84 mmol, 14 equivalents) of 2,2',2''-[10-(1-{[2-(4-nitrophenoxy)-2-oxoethyl]amino}-1-oxopropane- 2-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl]triacetato gadolinium (see WO2001051095(A2)), followed by the resulting reaction mixture Was stirred and heated at 50° C. for 8 hours. The cooled solution was finally concentrated under reduced pressure to a volume of 15-20 mL. The concentrate was poured into 400 mL of ethyl acetate under stirring, the precipitate formed was filtered off and dried in vacuo. The solid was dissolved in water, the resulting solution was ultrafiltered with water using a 1 kDa membrane and the final concentrated water was lyophilized. The crude product was purified by RP chromatography to give 28 mg (10.6 μmol, 5%) of the title compound.
UPLC (ACN-HCOOH): Rt. = 0.41 min.
MS (ES ⁺ ): m/z (z=2)=11311.7 (M+2H) ²⁺ , m/z (z=3)=873.1 (M+3H) ³⁺ .

Example 8
2, 2', 2'', 2''', 2'''', 2''''', 2'''''', 2''''''', 2'''''''',2''''''''',2'''''''''',2''''''''''',2'''''''''''',2''''''''''''',2'''''''''''''',2''''''''''''''',2'''''''''''''''',2'''''''''''''''''-{3,7,10-triazatricyclo [3.3.3.0 ^1,5 ]undecane-3,7,10-triyltris[carbonyl (3,6,11,14-tetraoxo-4,7,10,13-tetraazahexadecane-8,2, 15-triyl)di-1,4,7,10-tetraazacyclododecane-10,1,4,7-tetrayl]} octadecaacetate hexagadolinium

Example 8a
Tetrahydro-1H,4H-3a,6a-(methanoiminomethano)pyrrolo[3,4-c]pyrrole
4.0 g (6.5 mmol) of 2,5,8-tris((4-methylphenyl)sulfonyl)tetrahydro-1H,4H-3a,6a-(methanoiminomethano)pyrrolo[3,4-c]pyrrole( Prepared by the procedure outlined in J. Org. Chem. 1996, 61, 8897-8903.) was refluxed in 44 mL aqueous hydrobromic acid (47%) and 24 mL acetic acid for 18 hours. The solvent was removed in vacuo, the residue was dissolved in water and the aqueous phase was washed twice with dichloromethane. The aqueous phase was lyophilized, taken up in a small amount of water and passed through an anion exchange column (DOWEX 1X8) eluting with water. The basic fractions were collected and concentrated to give 0.89 g of tetrahydro-1H,4H-3a,6a-(methanoiminomethano)pyrrolo[3,4-c]pyrrole as the free base.
¹ H-NMR (400 MHz, D ₂ O): δ=2.74 (s, 12 H) ppm.

Example 8b
tert-Butyl-{1-[5,8-bis{2,3-bis[(tert-butoxycarbonyl)amino]propanoyl}dihydro-1H,4H-3a,6a-(methanoiminomethano)pyrrolo[3,4 -C]Pyrrole-2(3H)-yl]-3-[(tert-butoxycarbonyl)amino]-1-oxopropan-2-yl}carbamate
431.5 mg (0.89 mmol, CAS [201472-68-6]) N-(tert-butoxycarbonyl)-3-[(tert-butoxycarbonyl)amino]alanine N,N-dicyclohexyl ammonium salt, 0.44 mL A solution in 4.3 mL DMF prepared from (2.54 mmol) N,N-diisopropylethylamine and 386 mg (1.0 mmol) HATU was added to 38.9 mg (254 μmol) tetrahydro-1H,4H in 2 mL DMF. Added to −3a,6a-(methanoiminomethano)pyrrolo[3,4-c]pyrrole. After stirring the combined mixture at room temperature for 20 minutes, the solvent was removed in vacuo and the residue was amino phase silica gel (ethyl acetate in hexane, 0 to 100%) followed by preparative HPLC (C18-Chromatorex 10 μm, Purification by chromatography with acetonitrile in water + 0.1% formic acid, 65% to 100%) and 68.6 mg of tert-butyl-{1-[5,8-bis{2,3-bis[(tert-butoxy Carbonyl)amino]propanoyl}dihydro-1H,4H-3a,6a-(methanoiminomethano)pyrrolo[3,4-c]pyrrole-2(3H)-yl]-3-[(tert-butoxycarbonyl)amino] 1-oxopropan-2-yl}carbamate was obtained.
¹ H-NMR (300 MHz, CDCl ₃ ): δ=1.43 s, br, 54 H), 3.34-3.97 (m, 18 H), 4.48 (s, br, 3 H), 5. 01-5.67 (m, 6H) ppm.
UPLC (ACN-HCOOH): Rt. = 1.48 min.
MS (ES ⁺ ): m/z = 1012.6 (M + H) ⁺ .

Example 8c
3,3′,3″-[1H,4H-3a,6a-(methanoiminomethano)pyrrolo[3,4-c]pyrrole-2,5,8(3H,6H)-triyl]tris(3- Oxopropane-1,2-diaminium)hexachloride
65 mg (60 μmol) of tert-butyl-{1-[5,8-bis{2,3-bis[(tert-butoxycarbonyl)amino]propanoyl}dihydro-1H,4H-3a,6a-(methanoiminomethano) Pyrrolo[3,4-c]pyrrole-2(3H)-yl]-3-[(tert-butoxycarbonyl)amino]-1-oxopropan-2-yl}carbamate (Example 8b) in 2.0 mL. Dissolved in DMF and added 0.48 mL hydrochloric acid in dioxane (4M, 0.19 mmol). The reaction mixture was heated under microwave irradiation with stirring at 80° C. for 10 minutes. The solvent was removed in vacuo, the residue was taken up in a little water, lyophilized to give 38.9 mg of 3,3',3"-[1H,4H-3a,6a-(methanoiminomethano)pyrrolo. [3,4-c]pyrrole-2,5,8(3H,6H)-triyl]tris(3-oxopropane-1,2-diaminium)hexachloride was obtained.
¹ H-NMR (600 MHz, D ₂ O): δ=3.40-3.50 (m, 3H), 3.52-3.56 (m, 3H), 3.79-4.19 (m , 12H), 4.51-4.54 (m, 3H) ppm.
UPLC (ACN-HCOOH): Rt. = 0.20 min.
MS (ES ⁺ ): m/z=412.3 ([M+H) ⁺ , free base.

Example 8
2, 2', 2'', 2''', 2'''', 2''''', 2'''''', 2''''''', 2'''''''',2''''''''',2'''''''''',2''''''''''',2'''''''''''',2''''''''''''',2'''''''''''''',2''''''''''''''',2'''''''''''''''',2'''''''''''''''''-{3,7,10-triazatricyclo [3.3.3.0 ^1,5 ]undecane-3,7,10-triyltris[carbonyl (3,6,11,14-tetraoxo-4,7,10,13-tetraazahexadecane-8,2, 15-triyl)di-1,4,7,10-tetraazacyclododecane-10,1,4,7-tetrayl]} octadecaacetate hexagadolinium
30 mg (48 μmol) of 3,3′,3″-[1H,4H-3a,6a-(methanoiminomethano)pyrrolo[3,4-c]pyrrole-2,5,8(3H,6H)-triyl ] Tris(3-oxopropane-1,2-diaminium)hexachloride (Example 8c) was dissolved in a mixture of 1.8 mL DMSO, 1.8 mL DMF and 116 μL pyridine. At 60°C, 281 mg (0.38 mmol, WO2001051095(A2)) of 2,2',2''-[10-(1-{[2-(4-nitrophenoxy)-2-oxoethyl]]. Amino}-1-oxopropan-2-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl]triacetato gadolinium, followed by 44 μL of trimethylamine to give The reaction mixture was stirred at 60° C. for 15 hours and at room temperature for 2 days. Another amount of 2,2′,2″-[10-(1-{[2-(4-nitrophenoxy)-2-oxoethyl]amino}-1-oxopropan-2-yl)-1,4 ,7,10-Tetraazacyclododecane-1,4,7-triyl]triacetato gadolinium (56 mg, 75 μmol) and trimethylamine (5.4 μL) were added at 60° C. and stirring was continued at 60° C. for 15 hours. The solvent was removed in vacuo, the residue was taken up in 200 mL water and the resulting solution was ultrafiltered using a 1 kDa membrane. After further concentrating the concentrated water twice with 200 mL of deionized water and continuing the ultrafiltration, the final concentrated water was freeze-dried. The residue was dissolved in a mixture of 1.6 mL DMSO, 1.6 mL DMF and 105 μL pyridine and 601 mg (0.35 mmol) 2,2',2''-[10-(1-{ [2-(4-Nitrophenoxy)-2-oxoethyl]amino}-1-oxopropan-2-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl]triaceter The third addition of togadolinium and 48 μL triethylamine was repeated. After stirring at 60° C. for 18 hours, the ultrafiltration procedure using a 1 kDa membrane was repeated and the concentrated water was lyophilized after filtering 3 times at 200 mL. The crude product was purified by preparative HPLC (XBrigde C18, 5 μm, acetonitrile in water +0.1% formic acid, 0% to 7%) to give 51 mg of the title compound.
UPLC (ACN-HCOOH long run): Rt. = 2.95 min.
MS(ES ⁺ ): m/z(z=3)=1360.4(M+3H) ³⁺ , m/z(z=4)=1021.3(M+4H) ⁴⁺ , m/z(z=5)=817 .5 (M+5H) ⁵⁺ .

Example 9
2, 2', 2'', 2''', 2'''', 2''''', 2'''''', 2''''''', 2'''''''',2''''''''',2'''''''''',2'''''''''''-(3,7,9-triazabicyclo [3.3.1] Nonane-3,7-diylbis{carbonyl-1,4-diazepan-6,1,4-triylbis[(2-oxoethane-2,1-diyl)-1,4,7,10 -Tetraazacyclododecane-10,1,4,7-tetrayl]}) dodecaacetate tetragadolinium

Example 9a
3,7,9-Triazabicyclo[3.3.1]nonane
220 mg (0.49 mmol) 3,9-dibenzyl-7-(phenylsulfonyl)-3,7,9-triazabicyclo[3.3.1]nonane (reviewed in Tetrahedron Letters, 2005, 46, 5577-5580). Was prepared according to the procedure described in Example 1) and was refluxed in 3.4 mL of aqueous hydrobromic acid (47%) and 1.8 mL of acetic acid for 17 hours. The solvent was removed in vacuo, the residue was dissolved in water and the aqueous phase was washed twice with dichloromethane. The aqueous phase was lyophilized, taken up in a small amount of water and passed through an anion exchange column (DOWEX 1X8) eluting with water. The basic fractions were collected and concentrated to give 29.6 mg of 3,7,9-triazabicyclo[3.3.1]nonane as the free base.
¹ H-NMR (400 MHz, D ₂ O): δ=2.88 (t, 2H), 3.15 (d, 8H) ppm.

Example 9b
6-(methoxycarbonyl)-1,4-diazepandiium dichloride
8.3 g (24.5 mmol) of methyl 1,4-dibenzyl-1,4-diazepan-6-carboxylate in 42 mL of methanol (prepared analogously to US Pat. No. 5,866,562, p. 9). To this was added 8.3 mL concentrated hydrochloric acid, 2 mL water and 830 mg palladium on charcoal (10%). The suspension was stirred under a hydrogen atmosphere at 40° C. for 5 hours and at room temperature for 17 hours. The mixture was filtered through a Celite® route, the filtrate was concentrated in vacuo, to which was added toluene twice and removed in vacuo. The residue was dissolved in water and freeze-dried to obtain 5.65 g of 6-(methoxycarbonyl)-1,4-diazepandiium dichloride.
¹ H-NMR (400 MHz, D ₂ O): δ=3.49-3.68 (m, 9H), 3.70-3.73 (m, 4H), 3.75 (s, 3H) ppm ．

Example 9c
Methyl 1,4-bis{[4,7,10-tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetyl}-1,4 -Diazepan-6-carboxylate
To 200 mg (0.78 mmol) 6-(methoxycarbonyl)-1,4-diazepandiium dichloride in 10 mL dichloromethane was added 10 mL (6.2 mmol) N,N-diisopropylethylamine and the mixture was stirred at room temperature for 5 times. Stir for minutes. 1.04 g (1.56 mmol) of tri-tert-butyl 2,2',2''-(10-{2-[(2,5-dioxopyrrolidin-1-yl)oxy]-2-oxoethyl} -1,4,7,10-Tetraazacyclododecane-1,4,7-triyl)triacetate (Cong Li et al., J. Am. Chem. Soc. 2006, 128, p. 15072-15073; S3-5 and Galibert et al., Bioorg. Med. Chem. Letters 2010 (20), 5422-5425) was added) and the mixture was stirred at room temperature for 18 hours. The solvent was removed under reduced pressure and the residue was purified by chromatography on amino phase silica gel (ethyl acetate in hexane, 20-100%, then ethanol in ethyl acetate 0-100%) to give 210 mg of the title compound. Obtained.
UPLC (ACN-HCOOH): Rt. = 0.94 min.
MS (ES ⁺ ): m/z = 1267.6 (M + 1H) ⁺

Example 9d
Dodeca-tert-butyl 2,2',2'',2''',2'''',2''''',2'''''',2''''''',2 '''''''', 2''''''''', 2'''''''''', 2'''''''''''-(3, 7, 9-triazabicyclo[3.3.1]nonane-3,7-diylbis{carbonyl-1,4-diazepan-6,1,4-triylbis[(2-oxoethane-2,1-diyl)-1, 4,7,10-Tetraazacyclododecane-10,1,4,7-tetrayl]}) dodecaacetate
305 mg (0.24 mmol) of methyl 1,4-bis{[4,7,10-tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl ] Acetyl}-1,4-diazepan-6-carboxylate (Example 9c) was dissolved in 3.9 mL THF and a solution of 6.6 mg lithium hydroxide in 0.87 mL water was added. After stirring for 15 minutes, the solvent was removed under reduced pressure and untreated 1,4-bis{[4,7,10-tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10 Lithium-tetraazacyclododecan-1-yl]acetyl}-1,4-diazepan-6-carboxylate (300 mg) was dissolved in 2.0 mL dichloromethane. 120 μL (0.71 mmol) N,N-diisopropylethylamine, 112 mg (0.30 mmol) HATU and 40 mg (0.30 mmol) 3H-[1,2,3]triazolo[4,5-b]pyridine-3 -Ol was added and stirred for 15 minutes, then a solution of 15 mg (0.12 mmol) 3,7,9-triazabicyclo[3.3.1]nonane in 1 mL dichloromethane was added and the mixture was stirred for 3 days. did. Another 170 mg untreated 1,4-bis{[4,7,10-tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecane in 1 mL dichloromethane In 1-yl]acetyl}-1,4-diazepan-6-carboxylate lithium, 67 mg (0.18 mmol) HATU, 24 mg (0.18 mmol) 3H-[1,2,3]triazolo[4, 5-b]Pyridin-3-ol was added over 15 minutes and 50 μL of N,N-diisopropylethylamine was added. After stirring for 15 minutes, the freshly prepared HATU solution was added to the reaction mixture. After 1 day, the solvent was removed under reduced pressure, toluene was added thereto 6 times, and the solvent was removed in vacuo. The residue was purified by chromatography on amino phase silica gel (ethyl acetate in hexane, 0-100%, then ethanol in ethyl acetate 0-40%) to give 181 mg of the title compound.
UPLC (ACN-HCOOH): Rt. = 0.78 to 0.84 min.
MS (ES ⁻ ): m/z (z=2)=1298.7 (M−2H) ²⁻

Example 9
2, 2', 2'', 2''', 2'''', 2''''', 2'''''', 2''''''', 2'''''''',2''''''''',2'''''''''',2'''''''''''-(3,7,9-triazabicyclo [3.3.1] Nonane-3,7-diylbis{carbonyl-1,4-diazepan-6,1,4-triylbis[(2-oxoethane-2,1-diyl)-1,4,7,10 -Tetraazacyclododecane-10,1,4,7-tetrayl]}) dodecaacetate tetragadolinium
390 mg (mmol) of dodeca-tert-butyl 2,2',2'',2''',2'''',2''''',2'''''',2''''''',2'''''''',2''''''''',2'''''''''',2'''''''''''- (3,7,9-triazabicyclo[3.3.1]nonane-3,7-diylbis{carbonyl-1,4-diazepan-6,1,4-triylbis[(2-oxoethane-2,1- Diyl)-1,4,7,10-tetraazacyclododecane-10,1,4,7-tetrayl]}) dodecaacetate (Example 9d) was dissolved in 10.8 mL of water and an aqueous hydrochloric acid solution (2M) was added. In addition, the solution was adjusted to pH 2.5. 440 mg (1.25 mmol) gadolinium(III) oxide were added and the mixture was stirred at 80° C. for 17 hours, during which the pH of the suspension changed to pH 5. The mixture was diluted with water, sonicated and filtered. The filtrate was ultrafiltered using a 1 kDa membrane. After further diluting the concentrated water with 100 mL of deionized water and continuing the ultrafiltration, the final concentrated water was freeze-dried. The crude product was purified by preparative HPLC (C18 YMC-ODS AQ, 10 μm, acetonitrile in water +0.1% formic acid, 1% to 10%) to give 14.5 mg of the title compound.
UPLC (ACN-HCOOH): Rt. = 0.34 min.
MS (ES ⁺ ): m/z (z = 2) = 1272.9 (M + 2H) 2 ⁺

Example 10
{4,10-bis(carboxylatomethyl)-7-[2-oxo-2-({3-({[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetra Azacyclododecan-1-yl]acetyl}amino)-2,2-bis[({[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane-1- Il]acetyl}amino)methyl]propyl}amino)ethyl]-1,4,7,10-tetraazacyclododecan-1-yl}acetate tetragadolinium

Example 10a
tert-Butyl {4,10-bis(2-tert-butoxy-2-oxoethyl)-7-[2-oxo-2-({3-({[4,7,10-tris(2-tert-butoxy -2-Oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetyl}amino)-2,2-bis[({[4,7,10-tris(2-tert-butoxy -2-Oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetyl}amino)methyl]propyl}amino)ethyl]-1,4,7,10-tetraazacyclododecane-1 -Il} Acetate
6.6 mg (49.8 μmol, 1 eq) of 2,2-bis(aminomethyl)propane-1,3-diamine (see W. Hayes et al., Tetrahedron 59 (2003), 7983-7996) in 7 mL DMSO. Dissolved. 77 mg (0.6 mmol, 12 eq) N,N-diisopropylethylamine and 400 mg (0.6 mmol, 12 eq) tri-tert-butyl 2,2',2''-(10-{2-[(2 ,5-Dioxopyrrolidin-1-yl)oxy]-2-oxoethyl}-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate (M. Galibert et al., Bioorg. Med. Chem. Letters 2010 (20), 5422-5425 and J. Am. Chem. Soc. 2006, 128, p. 15072-15073; S3-5), and the resulting reaction mixture was stirred and Heated at °C overnight. The cooled solution was concentrated under reduced pressure. The crude product was used in the next chemical step without further characterization.

Example 10b
{4,10-bis(carboxymethyl)-7-[2-oxo-2-({3-({[4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclo Dodecane-1-yl]acetyl}amino)-2,2-bis[({[4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetyl }Amino)methyl]propyl}amino)ethyl]-1,4,7,10-tetraazacyclododecan-1-yl}acetic acid
The crude tert-butyl {4,10-bis(2-tert-butoxy-2-oxoethyl)-7-[2-oxo-2-({3-({[4,7,10 -Tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetyl}amino)-2,2-bis[({[4,7,10 -Tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetyl}amino)methyl]propyl}amino)ethyl]-1,4,7, 10-Tetraazacyclododecan-1-yl}acetate was dissolved in 40 mL TFA. The resulting solution was stirred at room temperature overnight and concentrated under reduced pressure. The crude product was used in the next chemical step without further characterization.

Example 10
{4,10-bis(carboxylatomethyl)-7-[2-oxo-2-({3-({[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetra Azacyclododecan-1-yl]acetyl}amino)-2,2-bis[({[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane-1- Il]acetyl}amino)methyl]propyl}amino)ethyl]-1,4,7,10-tetraazacyclododecan-1-yl}acetate tetragadolinium The crude {4,10-bis(from Example 10b. Carboxymethyl)-7-[2-oxo-2-({3-({[4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetyl }Amino)-2,2-bis[({[4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetyl}amino)methyl]propyl} Amino)ethyl]-1,4,7,10-tetraazacyclododecan-1-yl}acetic acid was dissolved in 10 mL of water. After adding 326 mg of tris(acetato-κO) gadolinium tetrahydrate, the pH value of the resulting solution was adjusted to 3.5-4.5 by adding aqueous sodium hydroxide solution. The reaction mixture was heated at 70° C. under stirring overnight. The resulting solution was ultrafiltered with water using a 1 kDa membrane and the final concentrated water was lyophilized. The crude product was purified by RP chromatography to give 65 mg (28 μmol, 46%) of the title compound.
UPLC (ACN-HCOOH): Rt. = 0.40 min.
MS (ES ⁺ ): m/z (z = 2) = 1149.7 (M + 2H) 2 ⁺ , m/z (z = 3) = 766.0 (M + 3H) ^{3 +} .

Example 11
[4,10-bis(carboxylatomethyl)-7-{2,5,11,14-tetraoxo-15-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetra Azacyclododecan-1-yl]-8,8-bis({[({[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl] Acetyl}amino)acetyl]amino}methyl)-3,6,10,13-tetraazapentadeca-1-yl}-1,4,7,10-tetraazacyclododecan-1-yl]acetate tetragadolinium

Example 11a
tert-Butyl [4,10-bis(2-tert-butoxy-2-oxoethyl)-7-{2,5,11,14-tetraoxo-15-[4,7,10-tris(2-tert-butoxy -2-Oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]-8,8-bis({[({[4,7,10-tris(2-tert-butoxy-2 -Oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetyl}amino)acetyl]amino}methyl)-3,6,10,13-tetraazapentadec-1-yl}- 1,4,7,10-Tetraazacyclododecan-1-yl]acetate
2.99 g (4.75 mmol, 12 eq) of N-{[4,7,10-tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1- Il]acetyl}glycine (see M. Suchy et al., Org. Biomol. Chem. 2010, 8, 2560-2566) and 732 mg (5.70 mmol, 14.4 eq) of ethyldiisopropylamine in 40 mL of N,N-dimethyl. Dissolved in formamide. 2.17 g of 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU; 5.70 mmol, 14.4 equivalents) ) Was added, the reaction mixture was stirred at room temperature for 15 minutes. 100.1 mg (396 μmol, 1 equivalent) of 2,2-bis(ammoniomethyl)propane-1,3-diaminium tetrachloride (see W. Hayes et al., Tetrahedron 59 (2003), 7983-7996) and 982. 7 mg (7.60 mmol, 19.2 eq) ethyldiisopropylamine were added and the resulting reaction mixture was stirred at 50° C. overnight. The cooled solution was concentrated under reduced pressure. The crude product was used in the next chemical step without further characterization.

Example 11b
[4,10-bis(carboxymethyl)-7-{2,5,11,14-tetraoxo-15-[4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclo Dodecane-1-yl]-8,8-bis({[({[4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetyl}amino )Acetyl]amino}methyl)-3,6,10,13-tetraazapentadeca-1-yl}-1,4,7,10-tetraazacyclododecan-1-yl]acetic acid
The crude tert-butyl [4,10-bis(2-tert-butoxy-2-oxoethyl)-7-{2,5,11,14-tetraoxo-15-[4,7,10 from Example 11a. -Tris(2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]-8,8-bis({[({[4,7,10-tris (2-tert-butoxy-2-oxoethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetyl}amino)acetyl]amino}methyl)-3,6,10,13-tetraaza Pentadeca-1-yl}-1,4,7,10-tetraazacyclododecan-1-yl]acetate was dissolved in 125 mL TFA. The resulting solution was stirred at 70° C. for 2 hours, room temperature overnight and concentrated under reduced pressure. The oily product was dissolved in 200 mL water, isolated by freeze-drying and used in the next chemical step without further characterization.

Example 11
[4,10-bis(carboxylatomethyl)-7-{2,5,11,14-tetraoxo-15-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetra Azacyclododecan-1-yl]-8,8-bis({[({[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl] Acetyl}amino)acetyl]amino}methyl)-3,6,10,13-tetraazapentadeca-1-yl}-1,4,7,10-tetraazacyclododecan-1-yl]acetate tetragadolinium Implemented The crude [4,10-bis(carboxymethyl)-7-{2,5,11,14-tetraoxo-15-[4,7,10-tris(carboxymethyl)-1,4, 7,10-Tetraazacyclododecan-1-yl]-8,8-bis({[({[4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane- 1-yl]acetyl}amino)acetyl]amino}methyl)-3,6,10,13-tetraazapentadeca-1-yl}-1,4,7,10-tetraazacyclododecan-1-yl] Acetic acid was dissolved in 100 mL of water. After adding 2.89 g of tris(acetato-κO) gadolinium tetrahydrate, the pH value of the resulting solution was adjusted to 3.0-3.5 by adding aqueous sodium hydroxide solution. The reaction mixture was heated at 70° C. for 24 hours under stirring. The resulting solution was ultrafiltered with water using a 1 kDa membrane and the final concentrated water was lyophilized. The crude product was purified by RP chromatography to give 296 mg (120 μmol, 30%) of the title compound.
UPLC (ACN-HCOOH): Rt. = 0.41 min.
MS (ES ⁺ ): m/z (z=2)=1262.8 (M+2H) ²⁺ , m/z (z=3)=841.5 (M+3H) ³⁺ .

Reference compound 1
Gadovist® (Gadobutrol, Bayer AG (Leverkusen, Germany))

Reference compound 2
Magnevist ® (Gadopentetate dimeglumine, Bayer AG (Leverkusen, Germany))

Reference compound 3
Primovist® (gadoxetate disodium, Bayer AG (Leverkusen, Germany))

Reference compound 4
Gadomer-17 was synthesized as described in EP 0836485 (B1) (Example 1k).

Characterization of Example Compounds In Vitro and In Vivo

The examples were tested once or multiple times in selected assays. When tested more than once, data are reported as either mean or median, where:
-The mean value, also called the arithmetic mean value, is the sum of the values obtained divided by the number of tests, and-the median value is the median value of the group of values when arranged in ascending or descending order. is there. If the number of values in the dataset is odd, the median is the median value. If the number of values in the dataset is even, the median is the arithmetic mean of the two middle values.

Examples were synthesized once or multiple times. When synthesized more than once, the data from the assay is the mean or median calculated utilizing the data sets obtained from testing one or more synthetic batches.

Example A
Measurement of relaxivity at 1.4T
Relaxability measurements at 1.41T were performed using a MiniSpec mq60 spectrometer (Bruker Analytik, Karlsruhe, Germany) operating at a resonance frequency of 60 MHz and a temperature of 37°C. T ₁ relaxation times were measured using the standard inversion recovery (IR) method with relaxation delay fixed at least 5 times T ₁ . Varying inversion time (TI) was automatically calculated by the MiniSpec mq60 standard software (8 steps). T ₂ was measured using a Carr-Purcell-Meiboom-Gill (CPMG) pulse sequence, applying a relaxation delay of at least 5 times that of T ₁ .

Each relaxivity measurement was performed with three different Gd concentrations (three concentrations between 0.05 and 2 mM). The T ₁ and T ₂ relaxation times of the compounds 1-10 of the examples were measured in different media, for example water, fetal bovine serum (FBS, Sigma, F7524) and human plasma.

Human plasma preparation: For each experiment, fresh blood was drawn from volunteers using a 10 mL citrate tube (Sarstedt S-Monovette 02.1067.001, 10 mL, Citrate). The 10 mL citrate tube was carefully inverted 10 times to mix the blood and anticoagulant and centrifuged at 1811 g for 15 minutes at room temperature (Eppendorf, Centrifuge 5810R).

The relaxivity r _i (where i=1, 2) was calculated based on the measured relaxation rate R _i in water and plasma:
R _i =R _{i (0)} +r _i [C _Gd ]
(In the formula, R _{i (0)} represents the relaxation rate of each solvent, and C _Gd represents the concentration of the compound normalized to gadolinium.) The gadolinium concentration of the investigated solution was determined by inductively coupled plasma mass spectrometry (ICP -Verified by MS Agilent 7500a (Waldbronn, Germany).

The relaxivity values thus obtained are summarized in Table 1.

Measurement of relaxivity at 3.0T
Relaxation at 3.0T was measured using a whole-body 3.0T MRI scanner (Philips Intera, Philips Healthcare (Hamburg, Germany)) and knee coil (SENSE-Knee-8, Philips Healthcare (Hamburg, Germany)). It was carried out. Sample tubes (CryoTube™ vials, Thermo Scientific 1.8 mL, Roskilde, Denmark) were placed in three rows of four and five sample tubes in a plastic holder in a box filled with water. The temperature was adjusted to 37°C. The shortest possible echo time (TE) of 7.46 ms was used for the MRI sequence. The inversion time was chosen to optimize the sequence to measure the T ₁ value corresponding to the estimated T ₁ range of all relaxation times of the contrast agent including solution. The following inversion times (TI) were applied: 50, 100, 150, 200, 300, 500, 700, 1000, 1400, 2100, 3200 and 4500 ms. After registering the final echo, the sequence was run with a constant relaxation delay of 3.4 seconds (TR varying from 3450 to 7900 ms). For details of the fitting method, see Rohrer et al. (Invest. Radiol. 2005;40, 11:715-724). The experimental matrix for phantom measurement was 320×320.

Relaxability was evaluated using 3 different concentrations of each compound (3 concentrations between 0.05 and 2 mM).

The T ₁ relaxation times of the compounds 1-6 of the examples were measured in water and human plasma. Human plasma preparation: For each experiment, fresh blood was drawn from volunteers using a 10 mL citrate tube (Sarstedt S-Monovette 02.1067.001, 10 mL, Citrate). The 10 mL citrate tube was carefully inverted 10 times to mix the blood and anticoagulant and centrifuged at 1811 g for 15 minutes at room temperature (Eppendorf, Centrifuge 5810R).

The relaxivity r _i (where i=1, 2) was calculated based on the measured relaxation rate R _i in water and plasma:
R _i =R _{i (0)} +r _i [C _Gd ]
(In the formula, R _{i (0)} represents the relaxation rate of each solvent, and C _Gd represents the concentration of the compound normalized to gadolinium (Table 2).)

Example B
Pharmacokinetic parameters The pharmacokinetic parameters of the compound of Example 3 were determined in male rats (Han-Wistar, 220-230g, n=3). The compound was administered as a sterile aqueous solution (52.5 mmol Gd/L) by bolus administration to the tail vein of animals. The dose was 0.1 mmol Gd/kg. 1 minute, 3 minutes, 5 minutes, 10 minutes, 15 minutes, 30 minutes, 60 minutes, 90 minutes, 120 minutes, 240 minutes, 360 minutes, 480 minutes after injection Blood was sampled after and 1440 minutes and the Gd concentration was measured by inductively coupled plasma mass spectrometry (ICP-MS Agilent 7500a (Waldbronn, Germany)). Blood levels were divided by 0.625 (the plasma fraction of rat blood assuming a strict extracellular distribution) to convert to plasma concentration. As a control, 3 animals were similarly treated with Gadovist®, a low molecular weight contrast agent. The time course of plasma levels is shown in Figure 1.

The obtained data was applied to three compartment models (Phoenix-WinNonlin) to obtain the pharmacokinetic parameters shown in Table 3.

Example C
Excretion after 5 days and residual gadolinium concentration in organs Excretion and organ distribution in Example 3 were confirmed in male rats (Han-Wistar, 100 to 110 g, n=3). The compound was administered as a sterile aqueous solution (54 mmol Gd/L) by bolus administration to the tail vein of the animal. The dose was 0.1 mmol Gd/kg. Urine is collected for a period of 0 to 1 hour, 1 to 3 hours, 3 to 6 hours, 6 to 24 hours, 1 to 2 days and 2 to 5 days after infusion, and feces for 0 to 1 day, 1 after injection. Collected for periods of ~2 and 2-5 days. As a control, 3 animals were similarly treated with Gadovist®, a low molecular weight contrast agent. On day 7, animals were sacrificed and the following organs were excised: blood, liver, kidney, spleen, heart, lung, brain, mesenteric lymph nodes, muscle, skin, stomach, intestine, bone and bone marrow. The remaining carcass was lyophilized and ground into a fine powder. Gd concentrations in organs and carcasses were measured by ICP-MS (ICP-MS Agilent 7500a (Waldbronn, Germany)). The results of organ distribution for Example 3 and Reference Compound 1 (Gadovist®) are summarized in Table 4. Example 3 is rapidly excreted via the kidney. Three hours later, 95.8%±3.4% of the infused amount was found in urine, and 5 days later, 96.9%±3.7% was found in urine. About 1.4% ± 0.6% was excreted by feces. Seven days after infusion, less than 0.5% of the dose is present in the body. With the exception of the excretory organ, the kidney, individual organs contained less than 0.03% of the infused volume.

Example D
Chemical stability Examples 1, 2, 3 and 6 were separately dissolved in 10 mM Tris-HCl buffer (pH 7.4) at a final concentration of 5 mmol Gd/L. An aliquot was taken out, and the remaining colorless and transparent solution was autoclaved at 121°C for 20 minutes. After autoclaving, the solution remained clear and colorless. An aliquot taken out before and after the autoclave treatment was analyzed by HPLC-ICP-MS to confirm the integrity of the compound.

HPLC: column: Hypercarb 2.5 mm x 15 cm. Solvent A: 0.1% formic acid in water. Solvent B: acetonitrile. Gradient from 100% A to 5% A + 95% B in 10 minutes. Flow rate 1 ml/min. Detection by ICP-MS matched to ¹⁵⁸ Gd. The chromatograms showing the intensities of Gd detected were visually compared. No change was observed in the chromatogram before and after the autoclave treatment. The compound was stable during the autoclave procedure.

Example E
Release of gadolinium after addition of zinc and phosphate
A proton relaxation time measurement method for evaluating transmetallation for measuring the stability of MRI contrast agents is described by Laurent S. et al. (Invest. Radiol. 2001; 36, 2:115-122). This technique is based on a phosphate buffer (pH 7.00, 26 mmol/L, KH ₂ PO ₄ Merck) containing 2.5 mmol/L gadolinium complex and 2.5 mmol/L ZnCl ₂ Sigma-Aldrich (Munich, Germany). (Hesse, Germany)) based on measurements of changes in the paramagnetic longitudinal relaxation rate of water protons in water. 100 microliters of a 250 mmol/L solution of ZnCl ₂ was added to 10 mL of a buffer solution of paramagnetic complex (reference compounds 1 to 4 and Example 3). The mixture was agitated vigorously and 300 μL withdrawn at 0 min, 60 min, 120 min, 3 h, 4 h, 5 h, 24 h, 48 h and 72 h for relaxation timing studies. The measurements were carried out at 60 MHz and 37° C. using a MiniSpec mq60 spectrometer (Bruker Analytik (Karlsruhe, Germany)). The results of Example 3 are shown in FIG. 2 in comparison with Reference Compound 1 (Gadovist®), Reference Compound 2 (Magnevist®) and Reference Compound 3 (Primovist®). If the transmetallation of gadolinium is triggered by Zn ²⁺ ions in phosphate buffer solution, then the released free Gd ³⁺ will react with the free PO ₄ ³⁻ ions to produce GdPO ₄ . Due to the low solubility of GdPO ₄ , some of the gadolinium precipitates as a solid, beyond which it does not affect the longitudinal relaxation rate of water. A reduced proton relaxation rate would be observed with the less stable gadolinium chelate [linear contrast agent in Figure 2: Reference compounds 2 (Magnevist ®) and 3 (Primovist ®). )]]. The stability of Example 3 is comparable to the highly stable reference compound 1 (Gadovist®).

Example F
Gd complex stability in human plasma at 37°C, 15 days Examples 3 and 10 were separately dissolved in human plasma at 1 mmol Gd/L. As a reference for the released Gd ³⁺ , 0.1 mmol/L gadolinium chloride (GdCl ₃ ) was dissolved in human plasma. Plasma samples were incubated at 37° C. for 15 days in a 5% CO ₂ atmosphere to keep the pH at 7.4. Aliquots were taken at the beginning and end of the incubation. The amount of Gd ³⁺ released from the complex was measured by HPLC-ICP-MS. Column: Chelated Sepharose (HiTrap, 1 mL). Solvent A: 10 mM Bis-Tris-HCl (pH 6.0). Solvent B: 15 mM HNO3. Gradient: 100% A for 3 minutes, 100% B for 3-10 minutes. Flow rate 1 mL/min. Detection by ICP-MS matched to ¹⁵⁸ Gd. A chromatogram showing the intensity of Gd detected was evaluated by peak area analysis. The size of the peak of Gd ³⁺ eluting after changing from solvent A to solvent B was recorded. For both compounds, this peak, and hence the increase in Gd ³⁺ release, was below the limit of quantification (less than 0.1% of the total amount of gadolinium injected). Both Gd complexes are stable under physiological conditions.

Example G
Solubility in Water The solubility of compounds in water was measured at room temperature (20° C.) in 0.5 mL of buffer solution (10 mM Tris-HCl) in a microcentrifuge tube (Eppendorf, 2.0 mL safety lock cap). Solid compound was added stepwise to the buffer solution. The suspension was mixed using a shaker (Heidolph Reax 2000) and treated in an ultrasonic bath (Bandelin, Sonorex Super RK255H) for 5 minutes. The suspension was stored at room temperature (20° C.) overnight, and the final gadolinium concentration was measured by inductively coupled plasma mass spectrometry (ICP-MS). The results are summarized in Table 5.

Example H
Magnetic resonance angiography with enhanced contrast (CE-MRA)
The potential for significant dose reduction is 100 μmol gadolinium per kilogram body weight [100 μmol Gd/kg bw] (this is equivalent to the standard dose in humans) and a low dose using 30 μmol gadolinium per kilogram body weight. Shown by in-house comparison with the protocol. As a representative of approved gadolinium-based MRI contrast agents, Reference Compound 1 (Gadovist®) was used in both dose protocols (100 μmol Gd/kg bw and 30 μmol Gd/kg bw), Example 3 ( 30 μmol Gd/kg bw).

A contrast-enhanced magnetic resonance angiography study was performed on a clinical 1.5T scanner (Magnetom Avanto, Siemens Healthcare, Erlangen, Germany). A standard spine coil was used for data collection for optimal signal utilization. The test was performed using male New Zealand white rabbits (weight 2.5-2.9 kg, n=6, Charles River Kisslegg). Weight-adjusted intramuscular injection of a mixture (1+2) of xylazine hydrochloride (20 mg/mL, Rompun 2%, Bayer Vital GmbH, Leverkusen) and ketamine hydrochloride (100 mg/mL, Ketavet, Pfizer, Pharmacia GmbH, Berlin). All animals were initially anesthetized with 1 mL/kg body weight. Continuous infusion of an intubated animal (endotracheal tube, Rueschelit Super Safe Clear, cuff 3.0 mm, Willy Ruesch AG (Kernen, Germany)) with propofol 0.9 mg/kg/h (10 mg/mL, Propofol-Lipuro 1% , B. Braun Melsungen AG (Mersungen, Germany)). Continuous intravenous injection was performed using the MR infusion system (Continuum MR Infusion System, Medrad Europe BV, AE Beek, Germany). Tracheal breathing (SV 900C, Maquet (Rastatt, Germany)) was performed at 55% oxygen, 40 breaths/min and a tidal volume of 7 mL/body weight/min.

The anatomic course of the aorta was obtained based on coronal, axial and sagittal localizer sequences. Time to peak is determined by a small intravenous test bolus (0.25 mL/2.5-2.7 kg or 0.3 mL/2.8-2.9 kg bw, reference compound 1) and 3D FLASH sequence (test bolus sequence). : Repetition time: 36.4 msec, echo time 1.45 msec, flip angle: 30 degrees, spatial resolution: 1.0×0.8×17 mm). The angiographic 3D FLASH sequence was characterized by a repetition time of 3.24 ms, an echo time of 1.17 ms, a flip angle of 25 degrees and a slice thickness of 0.94 mm. A 141 x 300 mm field of view was combined with a 150 x 320 matrix to give a spatial resolution of 0.9 x 0.9 x 0.9 mm and a total acquisition time of 13 seconds per 3D block. The 3D FLASH sequence was performed once before and immediately after the injection of the contrast medium. The time interval for intra-individual comparisons between administrations of different contrast agents was 20-30 minutes (animal n=3).

An intra-individual comparison magnetic resonance angiography picture obtained in rabbits is shown in FIG. 3: (A) 30 μmol Gd/kg bw of reference compound 1 (Gadovist®); (B) 30 μmol Gd/kg bw Example 3 and (C) 100 μmol Gd/kg bw Reference Compound 1. The contrast enhancement of the low dose protocol with Example 3 (B) is comparable to that of the standard dose of Reference Compound 1 (C). Moreover, the image quality of the low dose protocol of Example 3 (B) is far superior to that of the reference compound 1 (A). Angiographic studies show the potential for significant dose reduction of Example 3.

Example J
Whole Body Imaging Classic gadolinium-based extracellular contrast agents exhibit rapid extracellular passive distribution throughout the body and are exclusively excreted via the kidney. The rapid extracellular extracellular distribution, for example, has the potential for classical imaging as angiography and imaging of the central nervous system, limbs, heart, head/face/neck, abdomen and chest. to enable. The comparability of the pharmacokinetic and characteristic behaviors of Reference Compound 1 (Gadovist®) and other ECCMs has been demonstrated, which indicates that all bodies commonly imaged in precise diagnosis of various diseases. It forms the basis for bridging the effect on the site (Tombach B et al., Eur Radiol 2002; 12(6):1550-1556). The described contrast-enhanced magnetic resonance studies show that the pharmacokinetic distribution and diagnostic capabilities of Example 3 were compared to reference compound 1 (Gadovist®) as a representative of approved gadolinium-based MRI contrast agents. Are comparing.

To show that Example 3 has the same mechanism of action, MRI signal intensity and Gd concentration over time were measured in various tissues. The study was performed on a clinical whole-body MRI with a body spine coil, abdominal flex coil, and cervical coil (1.5T Magnetom Avanto, Siemens Healthcare, Erlangen, Germany). The test was carried out using male New Zealand white rabbits (weight 2.3-3.0 kg, n=8, Charles River Kisslegg). Weight-adjusted intramuscular injection of a mixture (1+2) of xylazine hydrochloride (20 mg/mL, Rompun 2%, Bayer Vital GmbH, Leverkusen) and ketamine hydrochloride (100 mg/mL, Ketavet, Pfizer, Pharmacia GmbH, Berlin). All animals were initially anesthetized with 1 mL/kg body weight. Continuous infusion of an intubated animal (endotracheal tube, Rueschelit Super Safe Clear, cuff 3.0 mm, Willy Ruesch AG (Kernen, Germany)) with propofol 0.9 mg/kg/h (10 mg/mL, Propofol-Lipuro 1% , B. Braun Melsungen AG (Mersungen, Germany)). Continuous intravenous injection was performed using the MR infusion system (Continuum MR Infusion System, Medrad Europe BV, AE Beek, Germany). Tracheal breathing (SV 900C, Maquet (Rastatt, Germany)) was performed at 55% oxygen, 40 breaths/min and a tidal volume of 7 mL/body weight/min.

Dynamic MRI measurements up to 22 minutes after injection followed by quantitative signal analysis (Siemens Mean Curve software (SYNGO Task Card, Siemens Healthcare (Erlangen, Germany)), head and neck (brain, tongue, oral muscles, neck muscles), It was performed on 3 different areas of the abdomen (spleen, liver, blood) and pelvis (limb muscles), 3D T1 weighted Vibe sequence was used for 3 different slice groups (TR=4.74 ms, TE=2.38). , Flip = 10°, 1:29 min.) Dynamic measurements of three slice groups (head/neck: 1:29 min, abdomen: 0:49 min, pelvis: 1:16 min) were performed up to 22 minutes after injection: 1. Head/neck: Baseline, 1.4, 5.2, 8.9, 12.8, 16.5, 20.4 minutes, 2. Abdomen: Baseline, 0.5, 4.3. , 8.1, 11.9, 15.7, 19.5 minutes and 3. Pelvis: Baseline, 2.9, 6.7, 10.5, 14.4, 18.1, 22.0 minutes. Thirty minutes after injection, animals were sacrificed and the following tissue samples were measured for Gd concentration using inductively coupled plasma mass spectrometry (ICP-MS Agilent 7500a (Waldbronn, Germany)): blood, brain, Tongue, liver and limb muscles.Quantitative image quality assessment was performed at 30 minutes post injection by a combination of quantitative ICP-MS gadolinium concentration and MRI region of interest analysis.

Administration of contrast agents leads to increased signals in the vascular system and the extravascular and extracellular spaces of the body. The enhancement of the signal is based on the pharmacokinetic and physicochemical properties of the contrast agent. FIG. 4 shows representative images of the head and neck area before and 1.4 minutes after administration of Example 3 and Reference Compound 1. FIG. 5 shows representative abdominal images before and 0.5 minutes after administration of Example 3 and Reference Compound 1. FIG. 6 shows representative images of the pelvic area before and 0.5 minutes after administration of Example 3 and Reference Compound 1. All images show a clear signal enhancement in, for example, the heart, tongue, aorta, kidneys, liver, spleen, entire vasculature and muscle.

The signal-time curve shows the change in signal over time after administration of the contrast agent and represents the pharmacokinetics of the contrast agent in each tissue (Fig. 7). A rapid increase in signal intensity was observed after contrast injection in all tissues examined, followed by a continuous decrease in signal. The degree of these contrast enhancements is tissue-specific. However, no difference in contrast improvement over time was observed between Example 3 and Reference Compound 1. This shows the same pharmacokinetic properties, indicating that Example 3 is suitable for different body regions (Figure 7). The magnitude of the contrast enhancement depends on tissue properties, especially tissue perfusion, and physicochemical properties, especially relaxivity. As expected from the approximately two-fold relaxivity (see Example A), the contrast enhancement using Example 3 is greater than that of Reference Compound 1.

The amount of gadolinium in the tissue at 30 minutes after injection and the signal in MRI measurements performed at 19.5 minutes (abdomen) after injection, 20.4 minutes (head and neck) after injection and 22.0 minutes (pelvis) after injection. The relationship between gadolinium concentration and changes in MRI signals was investigated by comparing the changes. The respective data for Example 3 and Reference Compound 1 are shown in FIG. No linear correlation was observed between gadolinium concentrations in various tissues and changes in the respective MRI signals. This indicates that the effects of Example 3 and Reference Compound 1 are independent of the body area or tissue investigated. A slight deviation from this correlation was observed in the spleen, indicating a stronger enhancement of the MRI signal than would be expected from the tissue concentration of gadolinium. This was observed with both contrast agents, but is associated with a very high blood volume in the spleen compared to other organs and tissues. Therefore, the spleen loses most of its gadolinium concentration by exsanguination, resulting in inconsistent imaging in vivo and ex vivo quantification of gadolinium. The correlation of signal changes in all other tissues and organs with tissue gadolinium concentration, which represents their relaxivity, depends on the effect of the contrast agent used. An even greater slope was obtained in Example 3 (1.9) than Reference Compound 1 (1.0), which is in good agreement with the higher known relaxivity of Example 3 (Fig. 8). See also relaxivity data described in Example A).

Example K
Dynamic CT Diffusion Phantom Study As shown in Example A, Reference Compound 4 has relaxivity that is in the same range as the compounds of the invention. Following intravenous infusion, all clinically approved small monomers GBCA (dimeglumine gadopentetate, meglumine gadoterate, gadoteridol, gadodiamide, gadobutolol and gadoversetamide) are distributed in the blood and extravascular/extracellular spaces by passive distribution (Aime S et al., J Magn Reson Imaging. 2009; 30, 1259-1267). Reversible binding to HSA, or long protein-binding contrast agents, such as gadophosbecet trisodium, that have a long duration in the blood vessel due to large hydrodynamic size, such as Reference Compound 4, may cause passage through the vessel wall. Disturbed. Due to the rapid renal excretion of GBCA, rapid diffusion within the vessel wall is necessary for good imaging results.

The dynamic CT diffusion studies described compare the ability of Examples 1, 2, 3, 4, 5, 6 and reference compounds 1 and 4 to cross semipermeable membranes (20 kDa). A 128-row clinical CT machine (SOMATOM Definition, 128; Siemens Healthcare (Forchheim, Germany)) was used to monitor diffusion through the semipermeable membrane at 100 kV and 104 mA. A dialysis cassette (Slide-A-Lyser, 20,000 MWCO, volume 0.1-0.5 mL, Thermo Scientific (Roskilde, Denmark)) filled with a contrast medium is placed in fetal bovine serum solution (FBS, Sigma, F7524). 0 minutes, 1 minute, 2 minutes, 3 minutes, 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30 minutes, 45 minutes, 60 minutes, 2 hours after placing Single measurements were taken at 3, 5, 7, 22, 22, 24, 30, 46, and 48 hours. Images were reproduced with a slice thickness of 2.4 mm and a B30 convolution kernel. The concentration used in the dialysis cassette of Examples 1, 2, 3, 4, 5, 6 and the reference compounds 1 and 4 investigated was 20 mmol Gd/L.

The imaging results of all investigated examples and reference compounds 1 and 4 at 0 minutes and 48 hours after placing the cassette in FBS solution are shown in FIG. The region of interest was hand-drawn in one centrally located slice for each time point for image analysis (a representative measurement region is shown in Figure 9: Image 1A). Figure 10 shows the results of Hounsfield units (HU) of the analyzed region over time. Calculated diffusion half-lives for the investigated examples and reference compounds are summarized in Table 6.

FIG. 10 and calculated half-life data show that Examples 1-6, as well as Reference Compound 1 (Gadovist®), in contrast to Reference Compound 4, are able to cross semipermeable membranes. . Furthermore, the data for the compounds investigated show that, in contrast to other high relaxivity agents that have high protein binding or very slow turnover rates (eg reference compound 4), the compounds of the invention provide a timely barrier. It is shown to have a hydrodynamic size that can be overcome. These findings demonstrate, for example, the ability of the compounds of the invention to overcome barriers such as the endothelial wall of the vascular system, which is a requirement for whole body imaging.

Example L
Assessing Potential Side Effects None of the compounds of the Examples investigated showed undesired negative side effects in the animals after dosing. In addition, the non-specific activity of Example 3 was screened by commercial radioligand binding and enzyme assays (LeadProfilingScreen®, Eurofins Panlabs (Taipei, Taiwan)), but no significant findings were revealed. ..

Example M
MRI with enhanced contrast of brain tumors in rats
The potential for a significant dose reduction is due to the low dose protocol using 0.3 mmol gadolinium per kilogram body weight (300 μmol Gd/kg bw) and 0.1 mmol gadolinium per kilogram body weight (100 μmol Gd/kg bw). It was shown by the intra-individual comparison of As a representative of approved gadolinium-based MRI contrast agents, Reference Compound 1 (Gadovist®) was used at both dose protocols (0.3 mmol Gd/kg bw and 0.1 mmol Gd/kg bw), It was compared with Example 3 (0.1 mmol Gd/kg bw).

GS9L cell line (European Collection of Cell Cultures, Cancer Res 1990; 50:138-141; J Neurosurg 1971; 34:335), 10% fetal bovine serum (FBS, Sigma F75249) and 1% penicillin-streptomycin (10. The cells were grown in Dulbecco's modified Eagle medium (DMEM, GlutaMAX™, Ref: 31966-021, Gibco) supplemented with 000 units/mL, Gibco). The test was performed using male Fisher rats (F344, body weight 170-240 g, n=4, Charles River Kisslegg). Inoculation was weight-adjusted with a mixture (1+2) of xylazine hydrochloride (20 mg/mL, Rompun 2%, Bayer Vital GmbH, Leverkusen) and ketamine hydrochloride (100 mg/mL, Ketavet, Pfizer, Pharmacia GmbH, Berlin). It was carried out under anesthesia with ketamine/xylazine using 1 mL/kg body weight using intramuscular injection. For intracerebral orthotopic transplantation, anesthetized animals were fixed in a stereotaxic apparatus and 1.0E+06 GS9L cells suspended in a volume of 5 μl of medium were slowly injected into the brain using a Hamilton syringe.

A contrast-enhanced MRI study was performed with a clinical 1.5T scanner (Magnetom Avanto, Siemens Healthcare (Erlangen, Germany)). A rat head coil (coil and animal holder for rats, RAPID Biomedical GmbH) was used for data collection. Rats were anesthetized with a mixture of isoflurane (2.25%), oxygen gas (about 0.5 L/min) and nitrous oxide (flow rate about 1 L/min). MR imaging was performed by 3D turbo spin echo sequence (121 mm slice in 3D block, field of view: 80 mm (oversampling 33%), repetition time: 500 ms, echo time 19 ms, spatial resolution: 0.3×0. 3×1.0 mm). Animals were imaged on two consecutive days. On day 1, Reference Compound 1 (Gadovist®) and Example 3 were compared within individuals at the same dose of 0.1 mmol Gd/kg bw, which is equivalent to the standard human dose. On day 2, 0.3 mmol Gd/kg bw of Reference Compound 1 (Gadovist®), which is equivalent to three times the human dose (clinically approved for certain CNS indications), is used in Example 3. It was compared with the standard dose (0.1 mmol Gd/kg bw). The resulting MR images of rat GS9L brain tumors are shown in Figure 11: an intra-individual comparison of Reference Compound 1 (Gadovist®) at the same 0.1 mmol Gd/kg body weight (bw) dose and Example 3 was performed. FIG. Example 3 showed higher lesion and brain contrast and excellent demarcation of tumor margin at the same dose. (B) Comparison of Example 3 with 0.3 mmol Gd/kg bw (triple dose) of reference compound 1 (Gadovist®) and 0.1 mmol Gd/kw bw (standard dose). Example 3 showed similar lesion and brain contrast at one third of the reference compound 1 dose.

Claims (15)

General formula (I)
(In the formula:
Is
Represents a group (in the group, * represents the point of attachment of the group to R ¹ );
R ¹ represents the group R ³ ;
n represents an integer of 4;
R ² represents a hydrogen atom;
R ³ is:
and
(In the group, * indicates the point of attachment of the group to the rest of the molecule)
Represents a group selected from:
R ⁴ represents a hydrogen atom;
R ⁵ represents a hydrogen atom or a methyl group)
Compound, or a stereoisomer, Oh Rui mixtures thereof. The compound according to claim 1, wherein R ⁵ represents a methyl group. R ³ is
The compound according to claim 1, which represents a group. R ³ is
The compound according to claim 1 or 2, which represents a group. [4,10-bis(carboxylatomethyl)-7-{3,6,12,15-tetraoxo-16-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetra Azacyclododecan-1-yl]-9,9-bis({[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane-1- Ill]propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetraazaheptadecane-2-yl}-1,4,7,10-tetraazacyclododecan-1-yl]acetate tetra gadolinium,
{4,10-bis(carboxylatomethyl)-7-[(2R,16R)-3,6,12,15-tetraoxo-16-[4,7,10-tris(carboxylatomethyl)-1,4 ,7,10-Tetraazacyclododecan-1-yl]-9,9-bis({[({(2R)-2-[4,7,10-tris(carboxylatomethyl)-1,4,7 , 10-Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetraazaheptadecane-2-yl]-1,4,7,10-tetra Azacyclododecan-1-yl}acetate tetragadolinium,
{4,10-bis(carboxylatomethyl)-7-[(2S,16S)-3,6,12,15-tetraoxo-16-[4,7,10-tris(carboxylatomethyl)-1,4 ,7,10-Tetraazacyclododecan-1-yl]-9,9-bis({[({(2S)-2-[4,7,10-tris(carboxylatomethyl)-1,4,7 , 10-Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetraazaheptadecane-2-yl]-1,4,7,10-tetra Azacyclododecan-1-yl}acetate tetragadolinium,
{4,10-bis(carboxylatomethyl)-7-[2-oxo-2-({3-({[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetra Azacyclododecan-1-yl]acetyl}amino)-2,2-bis[({[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane-1- Ill]acetyl}amino)methyl]propyl}amino)ethyl]-1,4,7,10-tetraazacyclododecan-1-yl}acetate tetragadolinium, and [4,10-bis(carboxylatomethyl)-7] -{2,5,11,14-tetraoxo-15-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl]-8,8- Bis({[({[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetyl}amino)acetyl]amino}methyl)-3, 6,10,13-Tetraazapentadeca-1-yl}-1,4,7,10-tetraazacyclododecan-1-yl]acetatotetragadolinium selected from the group consisting of: 3 or a compound according to any one of 4, or a stereoisomer, Oh Rui mixtures thereof. [4,10-bis(carboxylatomethyl)-7-{3,6,12,15-tetraoxo-16-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetra Azacyclododecan-1-yl]-9,9-bis({[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane-1- Ill]propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetraazaheptadecane-2-yl}-1,4,7,10-tetraazacyclododecan-1-yl]acetate tetra gadolinium:
In it, according to claim 1, 2 or a compound according to any one of 4, or a stereoisomer, Oh Rui mixtures thereof. Equation 4
(In the formula,
Is a tetraamine or a salt thereof as defined for the compound of general formula (I) according to any one of claims 1 to 6),
General formula (III):
Wherein R ⁵ is as defined for the compound of general formula (I) according to any one of claims 1 to 6 and LG represents an activated leaving group)
Of a compound of general formula (Id):
Where R ⁵ is as defined for the compound of general formula (I) according to any one of claims 1 to 4,
Is the tetraamine according to any one of claims 1 to 6. 7.) A method for preparing a compound of general formula (Id) according to any one of claims 1 to 6, comprising the step of providing a compound of
A composition comprising a compound according to any one of claims 1 to 6 for use in magnetic resonance imaging (MRI). Use of a compound according to any one of claims 1 to 6 or a mixture thereof for the manufacture of a diagnostic agent. Use of a compound or a mixture thereof according to any one of claims 1 to 6 for the preparation of a contrast agent for magnetic resonance imaging. Administering to a patient an effective amount of one or more compounds according to any one of claims 1 to 6 in a pharmaceutically acceptable carrier, and performing magnetic resonance imaging on the patient. A composition comprising the compound for imaging body tissue of a patient , comprising: General formula (III) for the preparation of a compound of general formula (I) according to any one of claims 1 to 6:
(Wherein R ⁵ is as defined for the compound of general formula (I) according to any one of claims 1 to 6 and LG represents an activated leaving group.)
Use of compounds. A compound of general formula (I′) for preparing a compound according to any one of claims 1, 2, 3, 4, 5 or 6.
(In the formula:
Is
Represents a group (in the group, * represents the point of attachment of the group to R ¹ );
R ¹ represents the group R ³ ;
n represents an integer of 4;
R ² represents a hydrogen atom;
R ³ is:
and
(In the group, * indicates the point of attachment of the group to the rest of the molecule)
Represents a group selected from:
R ⁴ represents a hydrogen atom;
R ⁵ represents a hydrogen atom or a methyl group)
Intermediate compound, or a stereoisomer, Oh Rui mixtures thereof. [4,10-bis(carboxylatomethyl)-7-{3,6,12,15-tetraoxo-16-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetra Azacyclododecan-1-yl]-9,9-bis({[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane-1- Ill]propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetraazaheptadecane-2-yl}-1,4,7,10-tetraazacyclododecan-1-yl]acetic acid,
{4,10-bis(carboxymethyl)-7-[(2R,16R)-3,6,12,15-tetraoxo-16-[4,7,10-tris(carboxymethyl)-1,4,7 , 10-Tetraazacyclododecan-1-yl]-9,9-bis({[({(2R)-2-[4,7,10-tris(carboxymethyl)-1,4,7,10- Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetraazaheptadecane-2-yl]-1,4,7,10-tetraazacyclododecane -1-yl}acetic acid,
{4,10-bis(carboxymethyl)-7-[(2S,16S)-3,6,12,15-tetraoxo-16-[4,7,10-tris(carboxymethyl)-1,4,7 , 10-Tetraazacyclododecan-1-yl]-9,9-bis({[({(2S)-2-[4,7,10-tris(carboxymethyl)-1,4,7,10- Tetraazacyclododecan-1-yl]propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetraazaheptadecane-2-yl]-1,4,7,10-tetraazacyclododecane -1-yl}acetic acid,
{4,10-bis(carboxymethyl)-7-[2-oxo-2-({3-({[4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclo Dodecane-1-yl]acetyl}amino)-2,2-bis[({[4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetyl }Amino)methyl]propyl}amino)ethyl]-1,4,7,10-tetraazacyclododecan-1-yl}acetic acid, and [4,10-bis(carboxymethyl)-7-{2,5, 11,14-Tetraoxo-15-[4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl]-8,8-bis({[({[ 4,7,10-Tris(carboxymethyl)-1,4,7,10-tetraazacyclododecan-1-yl]acetyl}amino)acetyl]amino}methyl)-3,6,10,13-tetraaza 14. The intermediate compound according to claim 13 selected from the group consisting of pentadeca-1-yl}-1,4,7,10-tetraazacyclododecan-1-yl]acetic acid, or a stereoisomer thereof. , Oh Rui mixtures thereof. [4,10-bis(carboxylatomethyl)-7-{3,6,12,15-tetraoxo-16-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetra Azacyclododecan-1-yl]-9,9-bis({[({2-[4,7,10-tris(carboxylatomethyl)-1,4,7,10-tetraazacyclododecane-1- Ill]propanoyl}amino)acetyl]amino}methyl)-4,7,11,14-tetraazaheptadecane-2-yl}-1,4,7,10-tetraazacyclododecan-1-yl]acetic acid:
In it, the intermediate compound according to claim 14, or a stereoisomer, Oh Rui mixtures thereof.